Substituted 4-thio pyridones

ABSTRACT

Novel cephem derivatives represented by the general formula ##STR1## in which the Acyl substituent is a group of the formula ##STR2## wherein Ar is a lipophilic optionally substituted phenyl, naphthyl or pyridyl group; R 1  represents either the residue of a heterocyclic amino acid or a C 2  -C 10  alkyl group substituted by both a carboxyl group and a group of the formula --NR 9  R 10  or ##STR3## in which R 9  and R 10  are each independently hydrogen or C 1  -C 6  alkyl, said C 2  -C 10  alkyl group being optionally interrupted by one or more nitrogen atoms or carbonyl groups, and R 2  and R 3  are each independently hydrogen, alkyl or aminoalkyl are gram-positive antibacterial agents, especially useful in the treatment of diseases caused by methicillin-resistant Staphylococcus aureus (also referred to below as MRSA or methicillin-resistant S. aureus).

CROSS-REFERENCE TO RELATED APPLICATION(S)

This is a division of application U.S. Ser. No. 08/427,778 filed Apr. 25, 1995, now U.S. Pat. No. 5,620,969.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention is directed to new cephem derivatives represented by the general formula ##STR4## in which the Acyl substituent is a group of the formula ##STR5## where Ar is an optionally substituted lipophilic phenyl, naphthyl or pyridyl group, R¹ represents either the residue of a heterocyclic amino acid or a C₂ -C₁₀ alkyl group substituted by both a carboxyl or sulfonyl group and a group of the formula --NR⁹ R¹⁰ or ##STR6## in which R⁹ and R¹⁰ are each independently hydrogen or C₁ -C₆ alkyl, said C₂ -C₁₀ alkyl group being optionally interrupted by one or more nitrogen atoms or carbonyl groups, and R² and R³ are each independently hydrogen, alkyl or aminoalkyl. The derivatives are gram-positive antibacterial agents, especially useful in the treatment of diseases caused by methicillin-resistant Staphylococcus aureus (also referred to below as MRSA or methicillin-resistant S. aureus).

2. Description of the Prior Art

The literature discloses a vast number of cephem derivatives having a wide variety of C-3 and C-7 substituents. Applicants are not aware, however, of any literature disclosing compounds with the combination of C-3 and C-7 substituents found by applicants to give good activity against MRSA organisms. There are, however, references which disclose cephalosporins having the type of 7-substituents or the type of 3-substituents present in the compounds claimed in the present application.

For example, there are references which disclose applicants' C-7 substituents. Among such references disclosing 7-substituents of the type ##STR7## where Ar is an aromatic group are the following:

U.S. Pat. No. 4,056,676 discloses cephem derivatives of the general formula ##STR8## where Z is hydrogen or fluorine; and when Z is hydrogen, each of X and Y is hydrogen or chlorine selected so that the phenyl ring is substituted with 1 or 2 chlorine atoms and so that when one chlorine atom is present said chlorine atom is in the 3-position, and when two chlorine atoms are present said chlorine atoms are in the 3,4-, the 3,5-or the 2,5-positions; and when Z is fluorine, said fluorine is in the 3- or 4-positions of the phenyl ring and each of X and Y is hydrogen or chlorine selected so that when the phenyl ring is substituted with 1 or 2 chlorine atoms, one of the chlorine atoms is in the 3- or 4-position of the phenyl ring; R¹ is hydrogen, dicyclohexylamine, or a pharmaceutically acceptable cation; and R is, inter alia, N-pyridino. Among the compounds specifically disclosed are those of the formulae: ##STR9## The compounds disclosed are said to be useful for treating and inhibiting the growth of MRSA organisms.

The cephalosporin derivative of the formula ##STR10## is disclosed in Antimicrobial Agents and Chemotherapy--1966, pg. 573-580 at page 576 (Compound No. 48).

J. Antibiotics, 26(12), 737-744, 1973, discloses the compound of the formula ##STR11##

U.K. Patent 998,265 discloses cephem derivatives of the general formula ##STR12## in which R¹, taken alone, is --OH, C_(1-C) ₈ acyloxy, or tertiaryamino, R² is -OH when R¹ is --OH, R² is --OH when R¹ is C₁ -C₈ acyloxy, R² is --O-- when R¹ is tertiaryamino, R¹ and R², when taken together, are --O--, R³ and R⁴ represent hydrogen, alkyl radicals having from 1 to 6 carbon atoms, alkenyl radicals having from 2 to 6 carbon atoms, cycloalkyl radicals having from 5 to 7 carbon atoms, or alkoxyalkyl radicals having from 2 to 6 carbon atoms; n represents 0 to 4; and R⁵ represents an alkyl radical having from 1 to 6 carbon atoms, an alkenyl or alkynyl radical having from 2 to 6 carbon atoms, a cycloalkyl radical having 5 or 6 carbon atoms, phenyl, β-furyl, β-thienyl, thienyl, or naphthyl, or a fluoro, chloro, bromo, nitro, trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄ alkylmercapto, or C₁ -C₄ alkoxy substitution product of such radicals.

U.K. Published Application No. 2,007,221 A discloses cephalosporin derivatives of the formula ##STR13## wherein Y is hydrogen, chlorine, bromine, C₁ -C₄ alkyl or C₁ -C₄ alkoxy; Z is a bond, oxygen or sulfur; W is hydrogen, methyl amino, hydroxy, SO₃ H or COOR₄ wherein R₄ is hydrogen or 5-indanyl with the proviso that when Z is oxygen or sulfur, W is other than hydroxy; R₁ is hydrogen or methoxy; R₂ is hydrogen, acetoxy, 1,3,4-thiadiazol-2-ylthio, 5-methyl-1,3,4-thiadiazol-2-ylthio, tetrazol-5-ylthio, 1-methyltetrazol-5-ylthio, 1,3,4-oxadiazol-2-ylthio, 5-methyl-l,3,4-oxadiazol-2-ylthio, 1,3,4-triazol-2-ylthio, 5-methyl-l,3,4-triazol-2-ylthio, 1,2,3-triazol-5-ylthio, pyridinium or 4-aminocarbonylpyridinium; R₃ is hydrogen, a negative charge when R₂ is pyridinium or 4-aminocarbonylpyridinium, a cation of an alkali metal or an alkaline earth metal, ammonium or organic ammonium cations, C₁ -C₄ alkyl, (C₂ -C₅ alkanoyloxy)methyl, (C₂ -C₅) alkanoylamino)methyl, [C₂ -C₅ alkanoyl(C₁ -C₄ alkoxy)carbonyl(C₁ -C₄ alkyl)-amino-methyl, p-(C₂ -alkanoyloxy)benzylamino(C₂ -C₁₅ alkanoyloxy)methyl, (C₁ -C₄ alkyl)amino(C₂ -C₁₅ alkanoyloxy)methyl or di(C₁ -C₄ alkyl)amino(C₂ -C₁₅ alkanoyloxy)methyl; and pharmaceutically acceptable salts thereof.

U.S. Pat. No. 3,217,000 discloses cephem derivatives of the formula ##STR14## wherein Thi is 2-thienyl or 3-thienyl and R is a substituent at the 3 or 4 position of the pyridino ring selected from the group consisting of cyano, carboxy, carbamyl, N-methylcarbamyl, carbo(C₁ -C₄ alkoxy), hydroxy and (C₁ -C₄)alkanoyl; and the salts thereof with pharmaceutically acceptable acids.

There is also literature disclosing cephalosporins having 3-substituents of the type ##STR15## where R is an optionally substituted aliphatic or aromatic group. Among such references are those of the following:

U.S. Pat. No. 4,758,557 discloses cephalosporin derivatives of the general formula ##STR16## wherein A represents an alkanoyloxy group having 2-5 carbon atoms; a carbamoyloxy group; an azido group; or an unsubstituted or substituted pyridylthio group of the formula ##STR17## where n is 0 or an integer of 3-5; R¹ and R² may be the same or different and each represents a hydrogen atom, a halogen atom, a carboxyl group or an optionally halogen-substituted lower-alkyl group having 1-5 carbon atoms; or an unsubstituted or substituted pyridiniumthio group of the formula ##STR18## where n, R¹ and R² have the same meanings as above; R³ represents a linear or branched-chain alkyl group having 1-5 carbon atoms, a halogen-substituted alkyl group, a cyclopropyl group, a cyclopropylmethyl group, an alkenyl group, an oxygen afore or a group of (--CH₂)_(m) --B where m is an integer of 0-3 and B represents a hydroxyl group, an alkoxy group, an amino group, an alkyl-substituted amino group, a carboxyl group, a carbamoyl group, a sulfonic acid group, a sulfonic acid amide group, a hydroxamic group, a cyano group, a thiol group, an alkylthio group, a methanesulfonylaminocarbonyl group or an acetamidosulfonyl group; or an unsubstituted or substituted pyridinium group of the formula ##STR19## where n has the same meaning as above; R⁴ and R⁵ may be the same or different and each represent a hydrogen atom, a linear or branched alkyl group having 1-5 carbon atoms, a carboxyl group, a carbamoyl group, a sulfonic acid group, a sulfonic acid amide group, a linear or branched alkylthio group having 1-5 carbon atoms, a halogen-substituted alkylthio group, a cycloalkanothio group, a carbamoylalkylthio group, an alkoxyalkylthio group or an alkyl-substituted aminoalkylthio group; or a 5- or 6-membered heterocyclicthio or bicycloheterocyclicthio group of the formula

    --S--Het

wherein Het represents an optionally substituted thiazole, isothiazole, 1,2,3-thiadiazole, 1,3,4-thiadiazole, 1,3,4-triazole, 1,2,3,4-tetrazole, pyrimidine, 1,2,4-triazine, benzothiazole, benzimidazole, benzoxazole, 1,3,4-triazaindolidine or 2,3-dihydro-1H-indolidinium group.

Illustrative of the compounds encompassed by U.S. Pat. No. 4,758,557 is MT0703 having the structure ##STR20## which is disclosed in J. Antibiotics 43(2), 189-198, 1990.

U.S. Pat. No. 4,786,633 discloses cephalosporin derivatives of the formula ##STR21## wherein R² is a substituted or unsubstituted heterocyclic group having 1-3 hetero atoms selected from the group consisting of nitrogen and sulfur. The R² group may be, for example, a group of the formula ##STR22## where R⁴ is lower alkenyl, lower alkyl, or a lower alkyl group substituted with a carbamoyl group and R⁵ and R⁶ are the same or different and each represent hydrogen or lower alkyl.

Published European Patent Application 409,164 A2 discloses cephalosporin derivatives of the formula ##STR23## wherein R¹ is an amino group or acylamino; R² is hydrogen or methoxy; R³ is hydrogen or a mono- or divalent substituent; R⁴ is optionally protected vic-dihydroxyaryl; R⁵ is straight or branched lower alkylene; R⁶ is hydrogen, a carboxy-protecting group or a negative charge when combined with Y; X is oxygen, sulfur, or sulfinyl; and Y is an anion or a negative charge when combined with R⁶ ; and the dotted line shows the presence or absence of a bond.

SUMMARY OF THE INVENTION

The present invention provides a novel series of cephem derivatives of the general formula ##STR24## wherein Ar is a group selected from ##STR25## in which R⁴, R⁵ and R⁶ are each independently hydrogen, halogen, trihalomethyl, nitro, amino, hydroxy, hydroxy(C₁ -C₆) alkyl, C₁ -C₆ alkyl, --(CH₂)_(n) OR⁷ or --(CH₂)_(n) SR⁷ ; n is an integer of from 1 to 6; R⁷ is hydrogen or C₁ -C₆ alkyl; R¹ is selected from the group consisting of

(a) a C₂ -C₁₀ alkyl group substituted by a carboxyl or sulfonyl group and a group of the formula ##STR26## in which R⁹ and R¹⁰ are each independently hydrogen or C₁ -C₆ alkyl, said C₂ -C₁₀ alkyl group being optionally interrupted by one or more nitrogen atoms or carbonyl groups; and

(b) a heterocyclic amino acid group of the formula ##STR27## R² and R³ are each independently hydrogen, C₁ -C₆ alkyl or amino(C₁ -C₆)alkyl; and R¹¹ is hydrogen, an anionic charge or a carboxyl-protecting group, provided that when R¹¹ is hydrogen or a protecting group, there is also present a counter ion; or a pharmaceutically acceptable salt or prodrug thereof. The compounds of formula I are antibacterial agents useful in the treatment of infections in humans and other animals caused by a variety of gram-positive bacteria, particularly methicillin-resistant S. aureus.

Also included in the invention are processes for preparing the compounds of formula I and pharmaceutical compositions containing said compounds in combination with pharmaceutically acceptable carriers or diluents.

DETAILED DESCRIPTION

The present invention provides novel cephem derivatives of general formula I above which are antibacterial agents useful in the treatment of infectious diseases in humans and other animals. The compounds exhibit good activity against a variety of gram-positive microorganisms, e.g. S. pneumoniae, S. pyrogenes, S. aureus, E. faecalis, E. faecium, S. epidermidis and S. hemolyticus, and are particularly useful against strains of methicillin-resistant S. aureus.

The compounds of formula I are characterized by a substituted pyridiniumthiomethyl group of the type ##STR28## at the 3-position of the cephem ring and a lipophilic 7-substituent of the type ##STR29## wherein Ar is an aromatic group selected from optionally substituted phenyl, naphthyl or pyridyl.

To elaborate on the definitions for the substituents of the formula I compounds:

(a) "Halogen" includes chloro, bromo, fluoro and iodo, and is preferably chloro or bromo;

(b) "Trihalomethyl" includes trichloromethyl, trifiuoromethyl, tribromomethyl and triiodomethyl, but is preferably trifiuoromethyl;

(c) The aliphatic "alkyl", "alkoxy", "alkenyl" and "alkynyl" groups may be straight or branched-chains having the specified number of carbon atoms, e.g., in the case of C₁ -C₁₀ alkyl, the alkyl group may have from 1 to 10 carbon atoms. It is preferred that the groups have up to 6 carbon atoms and most preferably up to 4 carbon atoms.

The term "pharmaceutically acceptable salt" as used herein is intended to include the nontoxic acid addition salts with inorganic or organic acids, e.g. salts with acids such as hydrochloric, phosphoric, sulfuric, maleic, acetic, citric, succinic, benzoic, fumaric, mandelic, p-toluenesulfonic, methanesulfonic, ascorbic, lactic, gluconic, trifluoro-acetic, hydroiodic, hydrobromic, and the like. Some of the compounds of the present invention have an acidic hydrogen and can, therefore, be converted with bases in a conventional manner into pharmaceutically acceptable salts. Such salts, e.g. ammonium, alkali metal salts, particularly sodium or potassium, alkaline earth metal salts, particularly calcium or magnesium, and salts with suitable organic bases such as lower alkylamines (methylamine, ethylamine, cyclohexylamine, and the like) or with substituted lower alkylamines (e.g. hydroxyl-substituted alkylamines such as diethanolamine, triethanolamine or tris- (hydroxymethyl) aminomethane), or with bases such as piperidine or morpholine, are also intended to be encompassed by the term "pharmaceutically acceptable salt".

Compounds of formula I in the form of acid addition salts may be written as ##STR30## where x.sup.⊖ represents the acid anion and R¹¹ is hydrogen or a carboxyl-protecting group. The counter anion x.sup.⊖ may be selected so as to provide pharmaceutically acceptable salts for therapeutic administration.

The carboxyl-protecting group R¹¹ is intended to include readily removable ester groups which have been employed to block a carboxyl group during the reaction steps used to prepare compounds I and which can be removed by methods which do not result in any appreciable destruction of the remaining portion of the molecule, e.g. by chemical or enzymatic hydrolysis, treatment with chemical reducing agents under mild conditions, irradiation with ultraviolet light or catalytic hydrogenation, etc. Examples of such protecting groups include benzhydryl, p-nitrobenzyl, 2-naphthhylmethyl, allyl, benzyl, p-methoxybenzyl, trichloroethyl, silyl such as trimethylsilyl, phenacyl, acetonyl, o-nitrobenzyl, 4-pyridylmethyl and C₁ -C₆ alkyl such as methyl, ethyl or t-butyl. Included within such protecting groups are those which are hydrolyzed under physiological conditions such as pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl, α-acetoxyethyl, α-pivaloyloxyethyl, and methoxymethyl. Compounds of formula I with such physiologically hydrolyzable carboxyl protecting groups are also referred to herein and in the claims as "prodrugs". Compounds of formula I where R¹¹ is a physiologically removable protecting group are useful directly as antibacterial agents. Compounds where an R¹¹ protecting group is not physiologically removable are useful intermediates which can be easily converted to the active form by conventional deblocking procedures well-known to those skilled in the art.

Compounds of formula I wherein a hydroxyl substituent is esterflied with a group hydrolyzable under physiological conditions are also included within the scope of the term "prodrug" as used herein and in the claims. Such hydroxyl protecting groups may be employed, for example, to increase the solubility of the formula I compound. Illustrative of suitable ester "prodrugs" of this type are compounds of formula I wherein one or more hydroxy substituent groups are converted to sulfate (--OSO₃ H) or phosphate (--OPO₃ H₂) groups.

A preferred embodiment of the present invention comprises compounds of formula I wherein Ar is ##STR31## in which R⁴, R⁵ and R⁶ are each independently hydrogen., halogen, C₁ -C₆ alkyl, trifluoromethyl, hydroxy, hydroxymethyl or amino.

Another preferred embodiment of the present invention comprises compounds of formula I wherein Ar is a group selected from ##STR32## in which R⁴, R⁵ and R⁶ are each independently hydrogen, halogen, trihalomethyl, nitro, amino, hydroxy, hydroxy(C₁ -C₆) alkyl, C₁ -C₆ alkyl, --(CH₂)_(n) OR⁷ or --(CH₂)_(n) SR⁷ ; n is an integer of from 1 to 6; R⁷ is hydrogen or C₁ -C₆ alkyl; R¹ is selected from the group consisting of ##STR33## wherein R⁸ is NR⁹ R¹⁰ or ##STR34## in which R⁹ and R¹⁰ are each independently hydrogen or C₁ -C₆ alkyl; m is 0 or 1; n is as defined above; o is 0 or an integer of from 1 to 4; p is 0 or 1; q is 0 or 1, with the proviso that q is 0 only when p is 0; r and s each represent an integer of from 1 to 4; R¹² is C₁ -C₆ alkyl; R² and R³ are each independently hydrogen, C₁ -C₆ alkyl or amino(C₁ -C₆)alkyl; and R¹¹ is hydrogen, an anionic charge or a carboxyl-protecting group, provided that when R¹¹ is hydrogen or a protecting group, there is also present a counter ion; or a pharmaceutically acceptable salt or prodrug thereof.

Within this group, the preferred compounds are those wherein Ar is ##STR35## in which R⁴, R⁵ and R⁶ are each independently hydrogen, halogen, C₁ -C₆ alkyl, trifluoromethyl, hydroxy, hydroxymethyl or amino.

The preferred individual compounds of the present invention, all of which have an MIC vs as a representative MRSA strain of ≦8 μg/ml, are listed below:

1-[(5S)-5-Amino-5-carboxy-1-pentyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 3)

1-[(5S)-5-Amino-5-carboxy-1-pentyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 4)

1-[(±)-5-Amino-5-carboxy-1-hexyl]4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio) acetamido]-5-thia-1 -azabicyclo [4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 5)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio) acetamido]-5-thia-1 -azabicyclo [4.2.0]-oct-2-en-3 -yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 6)

1-[(4S)4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,6-dichloropyridin-4-yl)thioacetamido]-5-thia1-azabicyclo [4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 7)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(3-bromophenylthio)acetamido]-5-thia1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 8)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(3,5-dichloro-4-hydroxyphenylthio)acetamido]-5-thia1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 9)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2-chloro-5-trifluoromethylphenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 10)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(3,5-dichloro-1-aminophenylthio)acetamido]-5-thia1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 11)

1-[(4S)4-Amino-4-carboxy-1-butyl]-4-[[( 6R)-trans-2-carboxy-8-oxo-7-[(2,5-dimethylphenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 12)

1-[(4S)-4-Amino-4-carboxy-1-butyl]4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,4,5-trichlorophenylthio)acetamido]-5-thia1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 13)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 14)

1-[(4S)-4-Amino4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,6-dichloropyridin-4-yl)thioacetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 15)

1-[(4S)4-Amino-4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichloro-4-hydroxymethylphenylthio)acetamido]-5 -thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 16)

1-[(4S)4-Amino-4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(3-fluorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 17)

1-[(4S)-4-Amino4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(3-bromophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 18)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2-chloro-5-trifluoromethylphenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 19)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,4,5-trichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 20)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2,3-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 21)

1-[(4S)-4-Amino4-carboxy-1-butyl]-2-ethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 22)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2-methyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 23)

1- [(4S) -4-Amino-4-carboxy-1-butyl]-2-(3-aminopropyl)-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 24)

1-[(4R)-4-Amino-4-carboxy-1 -butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio) acetamido]-5-thia-1-azabicyclo [4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 25)

1-[(4R)-4-Amino-4-carboxy-1-butyl]-4-[[(6 R) -trans-2-carboxy-8-oxo- 7-[(2,5-dichloro-4-hydroxymethylphenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 26)

1-[(4R)-4-Amino-4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 27)

1-[(3S)-3-Amino-3-carboxy-1-propyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 28)

1-[(2R)-2-Amino-2-carboxy-1-ethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 29)

1-[(2S)-5-Amino-2-carboxy-1-pentyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo [4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 30)

1-[(2S)-4-Amino-2-carboxy-1-butyl 4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 31)

1-[(2S)-4-Guanidino-2-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo [4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 32)

1-[(2S, 4R)-2-Carboxy-4-pyrrolidinyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 33)

1-[(1S)-1-(3-Amino-1-propyl)-N-(carboxymethyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 2)

1-[(1S)-1-(3-Guanidino-1-propyl)-N-(carboxymethyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 34)

1-[N-((1S)-1-carboxy-4-guanidino-1-butyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 35)

1-[(5S)-5-Amino-5-(N-carboxymethyl)carbamoyl-1-pentyl]-4-[[(6R) -trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 36)

1-[(4S)-4-Amino-4-(N-carboxymethyl)carbamoyl-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 37)

1-[N-(Carboxymethyl)-N-(2-amino-1-ethyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 38)

1-[N-(Carboxymethyl)-N-(3-amino-1-propyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio) acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 1)

1-[N-(Carboxymethyl)-N-(2-guanidino-1-ethyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 39)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(3-iodophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 40)

1-[(4S)-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(3-chlorophenylthio) acetamido]-5-thia-1-azabicyclo [4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 41)

1-[(4R)-4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo -7- [(3-iodophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 42)

1-[(4S)-4-Guanidino-4-carboxy-1-butyl]4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo [4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 43)

1-[(4S)-4-(t-Butoxycarbonylamino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(3-iodophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 44)

1-[(2S, 4S)-2-Carboxypyrrolidin-4-yl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo [4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 45)

The more preferred individual compounds of the present invention, all of which have a MIC≦8 μg/mL and a PD₅₀ <5 mg/kg against a representative strain of MRSA, are listed below:

1-[(5S) -5-Amino -5-carboxy-1-p entyl]-4- [[(6R) -trans-2-carboxy-8-oxo -7-[(2,5-dichlorophenylthio) acetamido]-5-thia-1-azabicyclo [4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 3)

1-[(5S)-5-Amino-5-carboxy-1-pentyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 4)

1-[(±)-5-Amino-5-carboxy-1-hexyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 5)

1-[(4S)4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 6)

1-[(4S)-4-Amino-4-carboxy-1-butyl]4-[[(6R) - trans-2-carboxy-8-oxo -7-[(2, 6 -dichloropyridin-4-yl) thioacetamido]-5-thia-1-azabicyclo[4-2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 7)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(3-bromophenylthio)acetamido]-5-thia-1-azabicyclo [4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 8)

1-[(4S)4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2-chloro-5-trifluoromethylphenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 10)

1-[(4S)4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(3,5-dichloro-4-aminophenylthio)acetamido]-5-thia-1-azabicyclo[4-2-0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 11)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 14)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,6-dichloropyridin-4-yl)thioacetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 15)

1-[(4S)-4-Amino4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichloro-4-hydroxymethylphenylthio)acetamido]--thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 16)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(3-bromophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 18)

1-[(4S)-4-Amino4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2-chloro-5-trifluoromethylphenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 19)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,4,5-trichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 20)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2,3-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 21)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2-ethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7- [(2,5-dichlorophenylthio) acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 22)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2-methyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 23)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2-(3-aminopropyl)-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 24)

1-[(4R)-4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 25)

1-[(4R)-4-Amino4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichloro-4-hydroxymethylphenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 26)

1-[(4R)-4-Amino-4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 27)

1-[(2S) -5-Amino-2-carboxy-1-pentyl]-4-[[(6R) -trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 30)

1-[(2S)-4-Amino-2-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 31)

1-[(2S)-4-Guanidino-2-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo [4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 32)

1-[(2S, 4R)-2-carboxypyrrolidin-4-yl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 33)

1-[(1S)-1-(3-Amino-1-propyl)-N-(carboxymethyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 2)

1-[(1S)-1-(3-Guanidino-1-propyl)-N-(carboxymethyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 34)

1-[N-((1S)-1-Carboxy-4-guanidino-1-butyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio) acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 35)

1-[(5S)-5-Amino-5-(N-carboxymethyl)carbamoyl-1-pentyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 36)

1-[(4S)-4-Amino-4-(N-carboxymethyl)carbamoyl-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 37)

1-[N-(Carboxymethyl)-N-(2-amino-1-ethyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 38)

1-[N-(Carboxymethyl)-N-(3-amino-1-propyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 1)

1-[(4S)-4-Guanidino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7[(2,5-dichlorophenylthioacetamido]-5-thia-1-azabicyclo [4.2.0]-oct-2-en-3 yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 43)

The most preferred individual compounds of the present invention are listed below:

1-[(4S)4-Amino-4-carboxy-1-butyl]4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 6)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,6-dichloropyridin-4yl) thioacetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 7)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(3-bromophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 8)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 14)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(3-bromophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable sale thereof (compound of Example 18)

1-[(4S)-4-Amino4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2-chloro-5-trifluoromethylphenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 19)

1-[(4S)4-Amino4-carboxy-1-butyl]-2,6-dimethyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,4,5-trichlorophenylthio) acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 20)

1-[(4S)-4-Amino-4-carboxy-1-butyl]-2,3-dimethyl-4-[[(6R)-(trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 21)

1-[(4S)-4-Amino4-carboxy-1-butyl]-2-methyl-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner sale or a pharmaceutically acceptable salt thereof (compound of Example 23)

1-[(4R)4-Amino-4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 25)

1-[(4R)-4-Amino4-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichloro-4-hydroxymethylphenylthio) acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 26)

1-[(2S)-4-Amino-2-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 31)

1-[(2S)-4-Guanidino-2-carboxy-1-butyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 32)

1-[(1S)-1-(3-Amino-1-propyl)-N-(carboxymethyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 2)

1-[N-((1S)-1-carboxy-4-guanidino-1-butyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 35)

1-[(5S)-5-Amino-5-(N-carboxymethyl)carbamoyl-1-pentyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 36)

1-[N-(Carboxymethyl)-N-(3-amino-1-propyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt or a pharmaceutically acceptable salt thereof (compound of Example 1)

The compounds of the present invention can be made by conventional methods. Two suitable procedures are summarized by the following reaction scheme: ##STR36##

To elaborate on the above process, thiol VII is converted into the arylthioacetic acid derivative VI by treatment with bromoacetic acid under basic conditions (e.g. aqueous sodium or potassium hydroxide). The reaction temperature for this step is typically between 20° C. and 100° C. Starting thiol VII is commercially available or can be prepared according to known literature procedures. Following acidification of the reaction mixture, the product VI is typically isolated by crystallization or, if necessary, it can be purified by chromatography.

Arylthioacetic acid VI is then coupled with a suitable cephem intermediate having a 3-substituent leaving group. For example, the leaving group may be acetoxy or halo. In the preferred embodiment illustrated by the reaction scheme, the cephem intermediate is the 3-chloro cephem V, but other suitable cephem intermediates with equivalent leaving groups at the 3-position could also be employed. The cephem intermediate V may be acylated with VI or a reactive derivative thereof by conventional acylation procedures well-known in the cephalosporin art to give N-acylated intermediate IV. In addition to using the free arylthioacetic acid, e.g. with a suitable condensing agent such as dicyclohexylcarbodiimide, acylating agent VI may also be employed in the form of equivalent acylating derivatives such as an acid anhydride, mixed anhydride, activated ester, or acid halide. The cephera intermediate preferably has the carboxyl group protected by a conventional carboxyl-protecting group which can be readily removed. Examples of such protecting groups are discussed above and include benzyl, 4-nitrobenzyl, 4-methoxybenzyl, diphenylmethyl, allyl, and the like. Other examples of suitable protecting groups are disclosed in Protective Groups in Organic Synthesis, Theodora W. Greene (John Wiley & Sons, 1981), Chapter 5. In one embodiment, intermediate V may be acylated with acid VI in the presence of dicyclohexylcarbodiimide and in an inert solvent such as tetrahydrofuran or dichloromethane. The reaction temperature is typically between -20° C. and 50° C. Upon completion of the reaction, insoluble material is removed by filtration, the filtrate is concentrated, and the residue is treated with a relatively non-polar solvent such as diethyl ether or ethyl acetate resulting in precipitation of the desired product. Alternatively, acid VI may be converted to the corresponding acid chloride, for example by treatment with thionyl chloride with or without a solvent such as dichloromethane, followed by coupling with cephem amine V in the presence of a base such as triethylamine or N-methylmorpholine to give intermediate IV. Cephem IV is typically isolated after aqueous work-up and evaporation of volatile solvents followed by trituration of the compound with a relatively non-polar solvent such as diethyl ether or ethyl acetate. This intermediate may be used in the next reaction step as the X=chloride derivative, or can be converted to the X=bromide or X=iodide derivative by treatment with the appropriate metal halide in a solvent such as acetone.

Conversion of cephem IV to the target quaternary cephems I is accomplished by two different methods. One method for preparation of I entails displacement of an appropriate 3-substituent leaving group with 4-mercaptopyridine followed by quaternization of the pyridyl nitrogen, and then deprotection of the cephera carboxylate ester. For example, cephem IV (X=Cl) is treated with optionally substituted 4-mercaptopyridine and sodium iodide in a one-pot reaction to give intermediate II. Reaction of thiopyridyl derivative II with a reactive alkylating agent provides the quaternary cephera intermediate I'. Examples of alkylating agents are α-halocarbonyl derivatives such as N-substituted haloacetamides. The alkylation reaction is carried out in an inert solvent such as acetone, dimethylformamide, or tetrahydrofuran and is run at temperatures between -20° C. and 100° C. Removal of the cephera carboxylate ester protecting group to give I is then accomplished under acidic conditions. For example, when R is diphenylmethyl or 4-methoxybenzyl, I is obtained upon treatment of I' with trifluoroacetic acid neat or in an inert solvent such as methylene chloride. A reagent such as anisole may also be employed to scavenge the liberated ester protecting group. The reaction temperature is usually at or below room temperature. The deprotection may also be carried out by treatment with other protic acids such as hydrochloric acid in a solvent such as methanol. The final product is typically isolated by precipitation or crystallization. In some cases, cephem I is purified by column chromatography, for example on reversed-phase adsorbent.

In a second method of preparing quaternary cephems I, intermediate IV is deprotected under acidic conditions, followed by reaction of the resulting intermediate IV' with a thiopyridone derivative III. For example, when R is diphenylmethyl or 4-methoxybenzyl, cephem acid IV'. is obtained upon treatment of IV with trifluoroacetic acid neat or in an inert solvent such as methylene chloride. A reagent such as anisole may also be employed to scavenge the liberated ester protecting group. The reaction temperature is usually at or below room temperature. The deprotection may also be carried out by treatment with other protic acids such as hydrochloric acid in a solvent such as methanol. The final product is typically isolated by precipitation or crystallization. Reaction of IV' with a thiopyridone derivative III in a solvent such as dimethylformamide, dimethyl sulfoxide, ethanol, methanol, or other appropriate solvents at a temperature between -20° C. and 100° C. affords target quaternary cephem I. The final product is isolated as described above. Thiopyridones III are typically prepared according to a method analogous to that described in T. Takahashi et al., European Patent Application No. 209751 and in I.E. El-Kholy et al., J. Heterocyclic Chem. Vol. 11, p. 487 (1974). This procedure entails reaction of 4-thiopyrone (European Patent No. 209751) with an appropriate primary amine in a solvent such as aqueous methanol or ethanol at a temperature ranging between 0° C. and 78° C. The primary amine may be in the form of a zwitterion in examples where there is a free acid group present in the molecule. In these cases, a base such as sodium hydroxide, sodium bicarbonate, or pyridine is added to form the free amine in situ. The product may be isolated as its sodium salt by evaporation of volatile solvents, followed by trituration with a solvent such as diethyl ether or ethyl acetate. Alternatively, the reaction mixture may be acidified and extracted with an organic solvent to afford the product as the free carboxylic acid. If the carboxylate group is protected as an ester, the amine may be free or present as an acid salt. In the latter case, a base such as sodium hydroxide, sodium bicarbonate, or pyridine is added to form the free amine in situ. The product is typically isolated by precipitation or by reversed phase column chromatography following removal of volatile solvents.

The thiopyridone derivatives of formula III are novel compounds and are another aspect of the present invention. The preferred R¹, R², and R³ substituents of derivatives III are as disclosed above in connection with the end-products of formula I.

It will be understood that where the substituent groups used in the above reactions contain certain reaction-sensitive functional groups such as amino or carboxylate groups which might result in undesirable side-reactions, such groups may be protected by conventional protecting groups known to those skilled in the art. For example, thiopyridone intermediates of formula III may have an amine functional group protected as the t-butyloxycarbamate. Suitable protecting groups and methods for their removal are illustrated, for example, in Protective Groups in Organic Synthesis, Theodora W. Greene (John Wiley & Sons, 1991). It is intended that such "protected" intermediates and end-products are included within the scope of the present disclosure and claims.

The desired end-product of formula I may be recovered either as the zwitterion or in the form of a pharmaceutically acceptable acid addition salt, e.g. by addition of the appropriate acid such as HCl, HI or methanesulfonic acid to the zwitterion. Compounds of formula I where R¹¹ is hydrogen or an anionic charge, or a pharmaceutically acceptable salt thereof, may be converted by conventional procedures to a corresponding compound where R¹¹ is a physiologically hydrolyzable ester group.

It will be appreciated that certain products within the scope of formula I may have a C-3 substituent group which can result in formation of optical isomers. It is intended that the present invention include within its scope all such optical isomers as well as epimeric mixtures thereof, i.e. R-- or S-- or racemic forms.

The novel cephalosporin derivatives of general formula I wherein R¹¹ is hydrogen, an anionic charge or a physiologically hydrolyzable carboxyl-protecting group, or the pharmaceutically acceptable salts or prodrugs thereof, are potent antibiotics active against many gram-positive bacteria. While they may be used, for example, as animal feed additives for promotion of growth, as preservatives for food, as bactericides in industrial applications, for example in waterbased paint and in the white water of paper mills to inhibit the growth of harmful bacteria, and as disinfectants for destroying or inhibiting the growth of harmful bacteria on medical and dental equipment, they are especially useful in the treatment of infectious disease in humans and other animals caused by the gram-positive bacteria sensitive to the new derivatives. Because of their excellent activity against MRSA organisms, they are particularly useful in the treatment of infections resulting from such bacteria.

The pharmaceutically active compounds of this invention may be used alone or formulated as pharmaceutical compositions comprising, in addition to the active cephera ingredient, a pharmaceutically acceptable carrier or diluent. The compounds may be administered by a variety of means, for example, orally, topically or parenterally (intravenous or intramuscular injection). The pharmaceutical compositions may be in solid form such as capsules, tablets, powders, etc. or in liquid form such as solutions, suspensions or emulsions. Compositions for injection, the preferred route of delivery, may be prepared in unit dose form in ampules or in multidose containers and may contain additives such as suspending, stabilizing and dispersing agents. The compositions may be in ready-to-use form or in powder form for reconstitution at the time of delivery with a suitable vehicle such as sterile water.

The dosage to be administered depends, to a large extent, on the particular compound being used, the particular composition formulated, the route of administration, the nature and condition of the host and the particular situs and organism being treated. Selection of the particular preferred dosage and route of application, then, is left to the discretion of the physician or veterinarian. In general however, the compounds may be administered parenterally or orally to mammalian hosts in an amount of from about 50 mg/day to about 20 g/day. Administration is generally carried out in divided doses, e.g., three to four times a day, analogous to dosing with a cephalosporin such as cefotaxime.

To illustrate the antibacterial properties of the compounds of the present invention, the following biological data is presented below.

IN VITRO ACTIVITY

Samples of the compounds prepared below in Examples 1-45 after solution in water and dilution with Nutrient Broth were found to exhibit the following ranges of Minimum Inhibitory Concentrations (MIC) versus the indicated microorganisms as determined by tube dilution. The MICs were determined using a broth micro dilution assay in accordance with that recommended by the National Committee for Clinical Laboratory Standards (NCCLS). Mueller-Hinton medium was used except for Streptococci which was tested in Todd Hewitt broth. The final bacterial inoculate contained approximately 5×10⁵ cfu/ml and the plates were incubated at 35° C. for 18 hours in ambient air (Streptococci in 5% CO₂). The MIC was defined as the lowest drug concentration that prevented visible growth.

    ______________________________________                                         Microorganism          MIC range in mcg/ml                                     ______________________________________                                         S. aureus methicillin resistant A27223                                                                 1-8                                                    S. pneumoniae A9585    0.001-0.06                                              S. pyogenes A9604      0.0005-0.06                                             E. faecalis A20688     0.125-4                                                 E. faecium A24885       0.5-16                                                 S. aureus A9537, penicillinase negative                                                               0.001-0.06                                              S. aureus A15090, penicillinase positive                                                              0.125-0.5                                               S. epidermidis A24548  0.007-0.25                                              S. epidermidis A25783, methicillin resistant                                                          0.06-0.5                                                S. hemolyticus A21638  0.015-0.25                                              S. hemolyticus A27235, methicillin resistant                                                          0.5-8                                                   ______________________________________                                    

IN VIVO ACTIVITY

The in vivo therapeutic efficacy of the compounds prepared in Examples 1-39 below after intramuscular injection to mice experimentally infected with the representative MRSA strain A27223 was also measured.

The Determination of the Effectiveness of Antimicrobial Agents in Staphylococcus aureus Systemic Infection in Mice

Organisms

The test organism, MRSA strain A27223 used to generate systemic infection in mice, is grown on two large Brain Heart Infusion Agar plates. On each plate, 0.5 ml of frozen stock culture is plated out. Plates are then incubated for 18 hours at 30° C. The next day each plate is washed with 20 ml of Brain Heart Infusion Broth and then pooled together. A microscopic direct count of microorganism is done using a 1:1000 dilution of plate wash. After a direct count is obtained, the number of organisms per milliliter is calculated. The count is adjusted to the desired amount of inoculum by diluting in 4% hog mucin. The desired challenge (amount of organisms given to mice) is 2.4×10⁸ cfu/0.5 ml/mouse for MRSA strain A27223. The mice are infected intraperitoneally with 0.5 ml of challenge. Ten non-treated infected mice are used as controls.

Mice

Mice used are male ICR mice. The average weight of the animals is from 20 to 26 grams.

Drug Preparation and Treatment

Compounds are tested at 4 dose levels, (25, 6.25, 1.56, and 0.39 mg/kg) and prepared in 5% cremophor, unless otherwise specified. Vancomycin is used as the control compound, and is dosed at 6.25, 1.56, 0.39, and 0.098 mg/kg. It is prepared in 0.1 M phosphate buffer. There are five infected mice per dose level, and they are treated with 0.2 ml of the test compound, preferably by intramuscular injection. Treatment begins 15 minutes and 2 hours post-infection.

Test Duration

A PD50 (the dose of drug given which protects 50% of mice from mortality) runs for 5 days. During this time, mortality of mice are checked every day and deaths are recorded. The cumulative mortality at each dose level is used to calculate a PD₅₀ value for each compound. Surviving mice are sacrificed at the end of day 5 by CO₂ inhalation.

Calculation

Actual calculation of PD₅₀ is performed with a computer program using the Spearman-Karber procedure.

Results

The in vivo efficacy, expressed as the PD₅₀ value, ranged from about 1 to 22 mg/kg (for certain compounds, more than one test was carried out; the indicated range is for at least one test result when multiple tests were done).

ILLUSTRATIVE EXAMPLES

The following examples illustrate the invention, but are not intended as a limitation thereof. The abbreviations used in the examples are conventional abbreviations well-known to those skilled in the art. Some of the abbreviations used are as follows:

h=hour(s)

tool=mole(s)

mmol=mmole(s)

g=gram(s)

THF=tetrahydrofuran

L=liter(s)

mL=milliliter(s)

Et₂ O=diethyl ether

EtOAc=ethyl acetate

MeOH=methanol

DMF=dimethylformamide

In the following examples, all temperatures are given in degrees Centigrade. Melting points were determined on an electrothermal apparatus and are not corrected. Proton and carbon-13 nuclear magnetic resonance (¹ H and ¹³ C NMR) spectra were recorded on a Bruker AM-300 or a Varian Gemini 300 spectrometer. All spectra were determined in CDCl₃, DMSO-d₆, CD₃ OD, or D₂ O unless otherwise indicated. Chemical shifts are reported in δ units relative to tetramethylsilane (TMS) or a reference solvent peak and interproton coupling constants are reported in Hertz (Hz). Splitting patterns are designated as follows: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; br, broad peak; dd, doublet of doublets; dt, doublet of triplets; and app d, apparent doublet, etc. Infrared spectra were determined on a Perkin-Elmer 1800 FT-IR spectrometer from 4000 cm⁻¹ to 400 cm⁻¹, calibrated to 1601 cm⁻¹ absorption of a polystyrene film, and are reported in reciprocal centimeters (cm⁻¹). Mass spectra were recorded on a Kratos MS-50 or a Finnegan 4500 instrument utilizing direct chemical ionization (DCI, isobutene), fast atom bombardment (FAB), or electron ion spray (ESI). Ultraviolet spectra were determined on a Hewlett Packard 8452 diode array spectrophotometer in the solvent indicated.

Analytical thin-layer chromatography (TLC) was carried out on precoated silica gel plates (60F-254) and visualized using UV light, iodine vapors, and/or staining by heating with methanolic phosphomolybdic acid. Column chromatography, also referred to as flash chromatography, was performed in a glass column using finely divided silica gel at pressures somewhat above atmospheric pressure with the indicated solvents. Reversed-phase analytical thin-layer chromatography was carried out on precoated reverse phase plates and visualized using UV light or iodine vapors. Reversed-phase column chromatography was performed in a glass column using Baker Octadecyl (C₁₈), 40 μm.

EXAMPLE 1

1-[N-(Carboxymethyl)-N-(3-amino-1-propyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt ##STR37## A. 2,5-Dichlorophenylthioacetic Acid

A mixture of 2,5-dichlorothiophenol (10.3 g, 57.5 mmol) and bromoacetic acid (8.03 g, 57.8 mmol) in water (225 mL) was treated with 10 N NaOH (13 mL, 130 mmol) and the mixture was heated at 100° C. for 1 h. The reaction mixture was then cooled to 0° C. and acidified to pH 1 with 6N HCl. The product precipitated and was collected by filtration to give 13.0 g (95% yield) of 2,5-dichlorophenylthioacetic acid as white crystals, m.p. 118° C. ¹ H NMR (300 MHz, CDCl₃) δ3.74 (s, 2 H), 7.15 (dd, J=2, 9 Hz, 1 H), 7.32 (d, J=9 Hz, 1 H), 7.36 (d, J=2 Hz, 1 H). Anal Calcd. for C₈ H₆ O₂ SCl₂ : C, 40.53; H, 2.55. Found: C, 40.46; H, 2.64.

B. (6R)-trans-3-Chloromethyl-7-[(2,5-dichlorophenyl)thioaceta-mido]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, diphenylmethyl ester

Method a

A solution of 2,5-dichlorophenylthioacetic acid (13.0 g, 54.9 mmol) in methylene chloride (55 mL) and thionyl chloride (10 mL, 137 mmol) was heated at reflux for 3 h. The reaction mixture was allowed to cool to room temperature and was concentrated in vacuo. The residue was evaporated two times from toluene to give 14 g of 2,5-dichlorophenylthioacetyl chloride (100% yield) as a slightly colored product which was used in the next step without purification. ¹ H NMR (300 MHz, CDCl₃)δ4.13 (s, 2 H), 7.22 (dd, J=2, 9 Hz, 1 H), 7.35 (d, J=9 Hz, 1 H), 7.39 (d, J=2 Hz, 1 H).

(6R) -trans-3-Chloromethyl-7-amino-8-oxo-5-thia-1-azabicyclo-[4.2.0]oct-2-ene-2-carboxylate, diphenylmethyl ester, HCl salt was stirred in a biphasic mixture of EtOAc and saturated NaHCO₃ for 0.5 h. The layers were separated, and the organic solution was dried over anhydrous MgSO₄, filtered, and concentrated to dryness. The free base (9.15 g, 22.0 retool) was dissolved in THF (200 mL), cooled to 0° C., and treated with N-methylmorpholine (3.34 g, 33.0 mmol) and 2,5-dichlorophenylthioacetyl chloride (6.75 g, 26.4 mmol). The reaction mixture was stirred for 1 h at 0° C., diluted with EtOAc (1000 mL) and washed with water (1000 mL) and brine (100 mL). The organic solution was then dried (MgSO₄) and the solvents were evaporated in vacuo. The residue was stirred with ether (100 mL). The product solidified and was collected by filtration to give 12.0 g (86% yield) of (6R)-trans-3-chloromethyl-7-[(2,5-dichlorophenyl)-thioacetamido]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, diphenylmethyl ester, m.p. 120° C. ¹ H NMR (300 MHz, CDCl₃)δ3.43 (d, J=18 Hz, 1 H), 3.59 (d, J=18 Hz, 1 H), 3.69 (d, J=17 Hz, 1 H), 3.79 (d, J=17 Hz, 1 H), 4.36 (d, J=12 Hz, 1 H), 4.41 (d, J=12 Hz, 1 H), 4.98 (d, J=5 Hz, 1 H), 5.81 (dd, J=5, 9 Hz, 1 H), 6.98 (s, 1 H), 7.14-7.44 (m, 14 H). Anal Calcd for C₂₉ H₂₃ N₂ O₄ S₂ Cl₃ : C, 54.94; H, 3.66; N, 4.42. Found: C, 55.18; H, 3.84; N, 4.62.

Method b

(6R)-trans-3-Chloromethyl-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, diphenylmethyl ester, HCl salt (Otsuka, 248 g, 0.55 mol) was treated with NaHCO₃ (56 g, 0.66 mol) in water (1.6 L) at 0° C. The mixture was stirred at 0° C. for 0.5 h and then CH₂ Cl₂ (1.5 L) was added. The biphasic mixture was filtered through Celite and the Celite pad was washed with CH₂ Cl₂ (2 L total). The layers were separated and the organic solution was dried over anhydrous MgSO₄, filtered, and concentrated to a volume of ca. 2 L. The free amine solution was then added to a mixture of 2,5-dichlorothio-phenylacetic acid (130 g, 0.55 mol) and dicyclohexylcarbodiimide (144 g, 0.70 mol) in THF (1 L) at room temperature. The reaction mixture was stirred for 2:5 h and then was filtered through Celite, washing the Celite pad with several portions of acetone. The filtrate was concentrated in vacuo to give a solid mass. The solid was slurried in Et₂ O and then collected by filtration, washing the solid with several portions of Et₂ O. The solid was dried under high vacuum over P₂ O₅ to give 268 g (77% yield) of (6R)-trans-3-chloromethyl-7-[(2,5-dichlorophenyl) thioacetamido]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, diphenylmethyl ester (see above for analytical data).

C. (6R)-trans-3-(4-Pyridylthiomethyl]-7-[(2,5-dichlorophenyl)-thioacetamido]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, diphenylmethyl ester

A solution of (6R)-trans-3-chloromethyl-7-[(2,5-dichlorophenyl)-thioacetamido]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, diphenylmethyl ester (6.00 g, 9.46 mmol) in acetone (100 mL) was treated with sodium iodide (4.26 g, 28.4 mmol). The mixture was stirred at 20° C. for 3 h and then condensed under reduced pressure to a volume of 50 mL. The concentrated solution was diluted with EtOAc (200 mL) and washed with ice water (100 mL). The organic solution was washed with saturated NaHSO₄ (20 mL), dried (MgSO₄), and evaporated under reduced pressure. The residue was stirred with ether (30 mL). The product solidified and was collected by filtration to give 6.40 g of (6R)-trans-3-iodomethyl-7-[(2,5-dichlorophenyl)thioaceta-mido]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, diphenylmethyl ester (93% yield) as a buff solid, m.p. 124° C. ¹ H NMR (300 MHz, CDCl₃)δ3.43 (d, J=18 Hz, 1 H), 3.69 (d, J=17 Hz, 1 H), 3.70 (d, J=18 Hz, 1 H), 3.78 (d, J=17 Hz, 1 H), 4.27 (d, J=9 Hz, 1 H), 4.33 (d, J=9 Hz, 1 H), 4.96 (d, J=5 Hz, 1 H), 5.75 (dd, J=5, 9 Hz, 1 H), 7.00(s, 1 H), 7.20-7.46 (m, 14 H). Anal. Calcd. for C₂₉ H₂₃ N₂ O₄ S₂ Cl₂ I: C, 48.01; H, 3.20; N, 3.86. Found: C, 48.00; H, 3.14; N, 3.76.

(6R)-trans-3-Iodomethyl-7-[(2,5-dichlorophenyl)thioacetamido]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, diphenylmethyl ester (3.00 g, 4.14 retool) was dissolved in THF (50 mL) at 0° C. and treated with 4-mercaptopyridine (0.504 g, 4.54 mmol). A solution of 2,6-lutidine (0.576 g, 5.38 mmol) in THF (1 mL) was added next, and the reaction mixture was stirred at 0° C. for 0.5 h and then at 20° C. for 1 h. The mixture was diluted with ethyl acetate (500 mL) and the organic solution was washed with water (2×500 mL) and brine (100 mL). The solution was then dried (MgSO₄) and evaporated under reduced pressure to give an oil which was treated with Et₂ O (50 mL) to give a solid. The solid was collected by filtration and purified by column chromatography on silica gel (CH₂ Cl₂ to 30% EtOAc/CH₂ C₂) to give 1.50 g of (6R)-trans-3-[(4-pyridylthiomethyl]-7-[(2,5-dichlorophenyl)-thioacetamido]-8-oxo-5-thia-1-azabicyclo [4.2.0]oct-2-ene-2-carboxylate, diphenylmethyl ester as a tan solid (49% yield), m.p. 122° C. ¹ H NMR (300 MHz, CDCl₃)δ3.37 (d, J=18 Hz, 1 H), 3.52 (d, J=18 Hz, 1 H), 3.68 (d, J=17 Hz, 1 H), 3.76 (d, J=17 Hz, 1 I-I), 3.96 (d, J=13 Hz, 1 H), 4.16 (d, J=13 Hz, 1 H), 4.93 (d, J=5 Hz, 1 H), 5.76 (dd, J=5, 9 Hz, 1 H), 6.95-7.42 (m, 16 H), 7.49 (d, J=9 Hz, 1 H), 8.29 (d, J=6 Hz, 2 H). Anal. Calcd. for C₃₄ H₂₇ N₃ O₄ S₃ Cl₂ : C, 57.62; H, 3.84; N, 5.93. Found: C, 57.27; H, 3.68; N, 5.79.

D. 1-[N-(p-Methoxybenzylcarboxymethyl)-N-(3-t-butyloxycarbonylamino-1-propyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium bromide

1. Preparation of N-(p-methoxybenzylcarboxymethyl)-N-[(3-t-butyloxycarbonylamino)-1-propyl]bromoacetamide

To a -78 ° C. solution of p-methoxybenzyl alcohol (6.91 g, 50 mmol) and triethylamine (6.95 mL, 50 mmol) in anhydrous CH₂ Cl₂ (50 mL) was added a solution of bromoacetyl bromide (10.1 g, 50 mmol) in CH₂ Cl₂ at -78° C. under nitrogen, over a period of 10 min. The cooling bath was removed, and the reaction mixture was stirred for 2 h. The mixture was washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to provide 12 g (93% yield) of p-methoxybenzyl bromoacetate as a dark colored oil which was used in the next reaction without purification. A portion was purified by column chromatography on silica gel (elution with 20% ethyl acetate/hexanes) to give a clear, colorless oil. ¹ H NMR (300 MHz, CDCl₃ ) δ3.79 (s, 3 H), 3.83 (s, 2 H), 5.12 (s, 2 H), 6.87 (m, 2 H), and 7.30 (m, 2 H); IR (film) 1738 cm⁻¹ ; MS (ESI/NH₄ OAc) M+NH₄ ⁺ =276.

To a cold (0-5° C.) solution of 3-(t-butyloxycarbonylamino)-1-propylamine (4.00 g, 23 mmol) and triethylamine (2.8 mL, 20 mmol) in anhydrous CH₂ Cl₂ (15 mL) was added slowly a solution of p-methoxybenzyl bromoacetate (5.18 g, 20 mmol) in anhydrous CH₂ Cl₂ (10 mL) over a period of 2.5 h. The mixture was stirred at 0°-5° C. for 2 h, at which time TLC indicated that the reaction was complete. The mixture was washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give a dark residue. The crude material was purified by column chromatography on silica gel (elution with 50% ethyl acetate/hexane) to afford 2.81 g (40% yield) of p-methoxybenzyl [3-(t-butyloxycarbonylamino)-1-propylamino]acetate as an amber oil. ¹ H NMR (300 MHz, CDCl₃ )δ1.41 (s, 9 H), 1.61 (quint, J=7 Hz, 2 H), 2.63 (t, J=7 Hz, 2 H), 3.17 (q, J=6 Hz, 2 H), 3.38 (s, 2 H), 3.78 (s, 3 H), 5.07 (s, 2 H), 6.86 (m, 2 H), and 7.26 (m, 2 H); IR (film), 1736, 1708 cm⁻¹ ; MS (isobutane-DCI) MH⁺ =353. Anal. Calcd. for C₁₈ H₂₈ N₂ O₅ : C, 61.35; H, 8.01; N, 7.95. Found: C, 60.87; H, 8.30; N, 7.89.

To a cold (0°-5° C.) solution of [3-(t-butyloxycarbonylamino)-1-propylamino]acetate (1.41 g, 4.0 mmol) and triethylamine (0.56 mL, 4.0 mmol) in anhydrous CH₂ Cl₂ (10 mL) was added over 5 mm a solution of bromoacetyl bromide (0.81 g, 4.0 mmol) in CH₂ Cl₂ under nitrogen. The reaction mixture was stirred at 0°-5° C. for 50 min and then diluted with CH₂ Cl₂, washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to give 1.81 g of an oily solid. Trituration with diethyl ether gave 1.35 g (71% yield) of N-(p-methoxybenzylcarboxymethyl)-N-[3-(t-butyloxy-carbonylamino)-1-propyl]bromoacetamide as white crystals, mp 79°-81° C. ¹ H NMR (300 MHz, CDCl₃)δ Two amide resonance structures: 1.40 and 1.42 (2 s, 9 H), 1.60 and 1.80 (2 quints, 2 H), 3.05 and 3.12 (2 m, 2 H), 3.43 (m, 2 H), 3.73 and 3.89 (2 s, 2 H), 3.79 and 3.80 (2 s, 3 H), 4.05 and 4.11 (2 s, 2 H), 4.60 and 5.28 (2 br s, 1 H), 5.08 and 5.12 (2 s, 2 H), 6.88 (m, 2 H), and 7.27 (m, 2 H); IR (KBr) 1736, 1684, and 1662 cm⁻¹ ; MS (ESI) MH⁺ =473. Anal. Calcd. for C₂₀ H₂₉ BrN₂ O₆ : C, 50.75; H, 6.17; N, 5.92. Found: C, 50.78; H, 6.22; N, 5.78.

2. Preparation of 1-[N-(p-Methoxybenzylcarboxymethyl)-N-(3-t-butyloxycarbonylamino-1-propyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium bromide

A solution of (6R)-trans-3-[(4-pyridylthiomethyl]-7-[(2,5-dichlorophenyl)-thioacetamido]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, diphenylmethyl ester (0.43 g, 0.60 mmol) in DMF (5 mL) was treated with N-(p-methoxybenzylcarboxymethyl)-N-[(3-t-butyloxycarbonylamino)-1-propyl]bromoacetamide (0.57 g, 1.2 mmol) at room temperature under nitrogen. After 96 h, the reaction mixture was added slowly to stirred diethyl ether (50 mL). A gum separated. The solvent was decanted and the gum was triturated with fresh diethyl ether to give a solid. The solid was collected by filtration, washed with additional ether, and dried to give 0.71 g (quantitative yield) of 1-[N-(p-methoxybenzylcarboxymethyl)-N-(3-t-butyloxy-carbonylamino-1-propyl)-carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)-acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium bromide. ¹ H NMR (DMSO-d₆, 300 MHz)δ1.39 (s, 9 H), 1.45-1.84 (m, 2 H), 2.81-3.07 (m, 2 H), 3.24-3.45 (m, 2 H), 3.55 (d, J=18 Hz, 1 H), 3.74 (s, 2 H), 3.82 (d, J=18 Hz, 1 H), 3.92 (s, 2 H), 4.13 (s, 1 H), 4.45 (s, 1 H), 5.05 (s, 2 H), 5.18 (d, J=5 Hz, 1 H), 5.45 (s, 1H), 5.53 (dd, J=5,8 Hz, 1 H),5.55 (s, 1 H), 6.84-6.95 (m, 4 H), 7.14-7.53 (m, 12 H), 7.92 (m, 2 H), 8.56 (m, 2 H), and 9.33 (d, J=8 Hz, 1 H); MS (ESI): M⁺ =1100.

E. 1-[N-(Carboxymethyl)-N-(3-amino-1-propyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo-[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium bromide, trifluoroacetate salt

A mixture of 1-[N-(p-methoxybenzylcarboxymethyl)-N-(3-t-butyloxycarbonylamino-1-propyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium bromide (0.70 g, 0.59 mmol) in CH₂ Cl₂ (6 mL) and anisole (0.6 mL) was treated with trifluoroacetic acid (6 mL) at room temperature. The resulting solution was stirred for 1 h and then concentrated in vacuo. Diethyl ether (30 mL) was added to give a nearly colorless precipitate. The solid was collected by filtration and dried to provide 0.47 g (87% yield) of 1-[N-(carboxymethyl)-N-(3-amino-1-propyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium bromide, trifluoroacetate salt as a light yellow solid. ¹ H NMR (DMSO-d₆, 300 MHz) δ1.63-2.05 (m, 2 H), 2.55-3.05 (m, 2 H), 3.26-3.66 (m, 2 H), 3.69 (d, J=18 Hz, 1 H), 3.80 (d, J=18 Hz, 1 H), 3.89 (s, 2 H), 4.00-4.38 (m, 2 H), 4.26-4.50 (m, 1 H), 5.12 (d, J=5 Hz, 1 H), 5.42 (s, 1 H), 5.60 (s, 1 H), 5.67 (dd, J=5,8 Hz, 1 H), 7.22 (dd, J=2,8 Hz, 1 H), 7.39-7.58 (m, 2 H), 8.00 (m, 2 H), 8.55 (m, 2 H), and 9.29 (d, J=8 Hz, 1 H); IR (KBr) 1776 cm⁻¹ ; MS (ESI) M⁺ =714. Anal. Calcd. for C₂₈ H₂₉ Cl₂ N₅ O₇ S₃.HBr.CF₃ CO₂ H: C, 39.61; H, 3.44; N, 7.70. Found: C, 39.72; H, 3.60; N, 7.48.

EXAMPLE 2

1-[(1S)-1-(3-Amino-1-propyl)-N-(carboxymethyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium inner salt ##STR38## A. (6R)-trans-3-Chloromethyl-7-[(2,5-dichlorophenyl)-thioaceta-mido]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid

A slurry of (6R)-trans-3-chloromethyl-7-[(2,5-dichlorophenyl)-thioacetamido]-8-oxo-5-thia-1-azabicyclo [4.2.0]oct-2-ene-2-carboxylate, diphenylmethyl ester (10.0 g, 15.8 mmol) in CH₂ Cl₂ (200 mL) at 0° C. was treated with anisole (24 mL) and then trifluoroacetic acid (80 mL). The resulting solution was stirred for 1 h at 0° C. and then concentrated under reduced pressure. The residue was stirred with Et₂ O, and the resulting solid was collected by filtration to give 5.20 g of (6R)-trans-3-chloromethyl-7-[(2,5-dichlorophenyl)-thioacetamido]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid as a white solid (70% yield), m.p. 125° C. ¹ H NMR (300 MHz, DMSO-d₆) 67 3.51 (d, J=18 Hz, 1 H), 3.70 (d, J=18 Hz, 1 H), 3.91 (s, 2 H), 4.52 (d, J=11 Hz, 1 H), 4.58 (d, J=11 Hz, 1 H), 5.13 (d, J=5 Hz, 1 H), 5.70 (dd, J=5, 8 Hz, 1 H), 7.24 (dd, J=2, 8 Hz, 1 H), 7.47 (dd, t=2, 8 Hz, 2 H), 9.28 (d, J=8 Hz, 1 H).

B. 1-[(1S)-1-(3-t-Butyloxycarbonylamino-1-propyl)-N-(t-butyl-carboxymethyl)-carbamoylmethyl]-4-[[(6R) -trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)-acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium chloride

1. Preparation of 1-[N-(t-butylcarboxymethyl)-1-[(t-butyloxycarbonyl-amino)propyl]-1-carbamoylmethyl]-4-thiopyridone

A solution of (2S)-[2-(benzyloxycarbonylamino)-5-(t-butyloxy-carbonylamino)]pentanoic acid (2.50 g, 6.82 mmol) in CH₂ Cl₂ (100 mL) was treated with N-hydroxysuccinimide (0.785 g, 6.82 mmol) followed by dicyclohexylcarbodiimide (1.41 g, 6.82 mmol). The mixture was stirred for 3 h and then treated with glycine t-butyl ester (0.895 g, 6.82 mmol, liberated from its HCl salt by dissolving in 1:1 dioxane/water and treating with 1 equiv. of sodium bicarbonate). After 18 h, the solvents were removed in vacuo, and the crude product was purified by flash chromatography on silica gel (elution with 5-10% MeOH/CH₂ Cl₂) to give 1.89 g (58% yield) of (2S)-N-t-butylcarboxy-methyl-2-(benzyloxycarbonylamino)-5-(t-butyloxycarbonyl-amino)pentanamide as a colorless oil. ¹ H NMR (300 MHz, CDCl₃ )δ1.42 (s, 9 H), 1.44 (s, 9 H), 1.56 (m, 2 H), 1.90 (m, 2 H), 3.09 (m, 1 H), 3.36 (br s, 1 H), 3.84 (dd, J=6, 21 Hz, 1 H), 3.98 (dd, J=6, 21 Hz, 1 H), 4.40 0br s, 1 H), 4.70 br s, 1 H), 5.11 (s, 2 H), 5.52 (d, J=6 Hz, 1 H), 6.88 (br s, 1 H), and 7.37 (m, 5 H).

A solution of (2S)-N-t-butylcarboxymethyl-2-(benzyloxy-carbonylamino)-5-(t-butyloxycarbonylamino)pentanamide (1.09 g, 2.27 mmol) in ethanol (30 mL) was treated with acetic acid (0.13 mL, 2.27 mmol). This solution was placed under a nitrogen atmosphere in a Parr bottle and then treated with 10% palladium on carbon (0.10 g, 0.09 mmol) and the resulting mixture was shaken in a Parr apparatus under 50 psi of hydrogen for 18 h. Filtration through Celite with the aid of 30 mL of water, followed by concentration of the filtrate in vacuo gave 0.903 g (98% yield) of the acetic acid salt of (25)-N-t-butylcarboxy-methyl-2-amino-5-(t-butyloxycarbonylamino)pentanamide as a colorless oil. ¹ H NMR (300 MHz, DMSO-d₆)δ1.35 (s, 9 H), 1.39 (s, 9 H), 1.39-1.71 (m, 4 H), 1.84 (s, 3 H, CH₃ CO₂ ⁻), 2.88 (q, J=6 Hz, 2 H), 3.16-3.20 (m, 1 H), 3.71 (d, J=4 Hz, 1 H), 3.72 (d, J=4 Hz, 1 H), 3.98 (br s, 3 H, NH₃ ⁺), 6.76 (t, J=5 Hz, 1 H), and 8.22 (t, J=6 Hz, 1 H); ¹³ C NMR (75 MHz, DMSO-d6) δ21.76, 25.83, 27.70, 28.27, 32.07, 33.35, 41.19, 54.05, 77.32, 80.57, 155.58, 168.96, 172.54, and 175.05; IR (film) 3320, 1739, 1690, 1534, 1368, 1162 cm⁻¹ ; MS (DCI) m/e 346 (MH⁺). Anal. Calcd. for C₁₆ H₃₁ N₃ O₅.C₂ H₄ O₂.0.25 H₂ O: C, 52.73; H, 8.73; N, 10.25. Found: C, 52.57; H, 8.94; N, 10.38.

A solution of the acetic acid salt of (2S)-N-t-butylcarboxymethyl-2-amino-5-(t-butyloxycarbonylamino)pentanamide (0.813 g, 2.00 mmol) in ethanol (10 mL) was treated with 1N NaOH (2.00 mL, 2.00 mmol). To this solution was added 4-thiopyrone (0.225 g, 2.00 mmol) (ref.: European Patent Application No. 209751). After stirring for 19 h, the reaction mixture was concentrated in vacuo to give 1.00 g of the crude product as a red oil. Purification by flash chromatography on silica gel (elution with 7.5% MeOH/CH₂ Cl₂) gave 0.498 g (57% yield) of 1-[(1S)-N-(t-butylcarboxymethyl)-1-[(t-butyloxycarbonylamino)propyl]]-carbamoylmethyl-4-thiopyridone as a red solid. ¹ H NMR (300 MHz, DMSO-d₆)δ1.35 (s, 9 H), 1.37 (s, 9 H), 1.58-1.72 (m, 2 H), 1.90-2.03 (m, 2 H), 2.88-2.96 (m, 2 H), 3.69-3.85 (m, 2 H), 4.75 (m, 1 H), 6.86 (t, J=6 Hz, 1 H), 7.15 (d, J=7 Hz, 2 H), 7.59 (d, J=7 Hz, 2 H), and 8.76 (t, J=6 Hz, 1 H); ¹³ C NMR (75 MHz, DMSO-d₆) δ24.47, 25.61, 27.67, 28.24, 28.53, 33.36, 41.59, 67.00, 77.55, 80.96, 129.91, 135.12, 155.61, 168.05, 168.29, and 190.46; IR (KBr) 3328, 1742, 1688, 1622, 1530, 1160, 1112 cm⁻¹ ; MS (ESI) m/e 440 (MH⁺). Anal. Calcd for C₂₁ H₃₃ N₃ O₅ S: C, 57.38; H, 7.57; N, 9.56. Found: C, 57.46; H, 7.82; N, 9.34.

2. Preparation of 1-[(1S)-1-(3-t-butyloxycarbonylamino-1-propyl)-N-(t-butylcarboxymethyl) carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium chloride

A solution of 1-[(1S)-N-(t-butylcarboxymethyl)-1-[(t-butyl-oxycarbonylamino)propyl]]carbamoylmethyl-4-thiopyridone (0.457 g, 1.04 mmol) in DMF (5 mL) was added to a solution of (6R)-trans-3-chloromethyl-7-[(2,5-dichlorophenyl)thioacetamido]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (0.489 g, 1.04 mmol) in DMF (5 mL). After 4 h, the reacton mixture was concentrated in vacuo to give 0.950 g of a red oil. Trituration with diethyl ether provided a solid which was collected by filtration to give 0.850 g of nearly pure material. The solid was stirred with ether for 2 h, and then collected by filtration and dried in vacuo to give 0.725 g (77% yield) of 1-[(1S)-1-(3-t-butyloxycarbonylamino-1-propyl)-N-(t-butylcarboxymethyl)carbamoyl-methyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium chloride as a tan solid. ¹ H NMR (300 MHz, DMSO-d₆)δ1.20 (m, 1 H), 1.34 (s, 9 H), 1.36 (s, 9 H), 1.54-1.67 (m, 1 H), 2.14 (m, 1 H), 2.25 (m, 1 H), 2.93 (m, 1 H), 3.52 (d, J=18 Hz, 1 H), 3.70-3.80 (m, 2 H), 3.92 (s, 2 H), 4.30-4.47 (m, 2 H, 5.14 (d, J=5 Hz, 1 H), 5.57 (t, J=7 Hz, 1 H), 5.67 (dd, J=5, 8 Hz, 1 H), 6.88 (m, 1 H), 7.23 (dd, J=2, 9 , Hz, 1 H), 7.44-7.48 (m, 2 H), 8.05 (d, J=7 Hz, 2 H), 8.83 (d, J=7 Hz, 2 H), and 9.30-9.32 (mm, 2 H); ¹³ C NMR (75 MHz, DMSO-d₆)δ25.44, 27.20, 27.68, 28.24, 28.96, 33.67, 34.12, 41.69, 47.49, 57.52, 59.38, 69.67, 77.59, 81.05, 122.53, 123.65, 126.22, 126.28, 126.94, 128.86, 130.69, 132.52, 137.89, 142.21, 155.59, 162.86, 163.32, 164.22, 167.05, 168.04, and 168.22; IR (KBr) 3328, 2978, 1782, 1686, 1626, 1542, 1450, 1368, 1160 cm⁻¹ ; MS (ESI) m/e 870 (MH⁺). Anal Calcd for C₃₇ H₄₅ N₅ O₉ S₃ Cl₂.HCl; C, 48.98; H, 5.11; N, 7.72. Found: C, 48.51; H, 5.11; N, 7.56.

C. 1-[(1S)-1-(3-Amino-1-propyl)-N-(carboxymethyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium chloride trifluoroacetate salt

A slurry of 1-[(1S)-1-(3-t-butyloxycarbonylamino-1-propyl)-N-(t-butylcarboxymethyl) carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo [4.2.0]-oct-2-en-3-yl]methylthio]pyridinium chloride (0.680 g, 0.750 mmol) in methylene chloride (20 mL) at 0° C. was treated with anisole (2.5 mL) followed by addition of trifluoroacetic acid (9.2 mL). The resulting solution was stirred at 0° C. for 30 min and then at room temperature for 5 h. The reaction mixture was concentrated in vacuo to give an oil. Trituration with ether gave a solid which was collected by filtration. The solid was slurried with acetone, collected by filtration, and dried in vacuo to give 0.219 g (33% yield) of 1-[(1S)-1-(3-amino-1-propyl)-N-(carboxymethyl)carbamoylmethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichlorophenylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]-oct-2-en-3-yl]methylthio]pyridinium chloride trifluoroacetate salt as a brown solid. ¹ H NMR (300 MHz, DMSO-d₆) δ1.32-1.57 (m, 2 H), 2.27 (br s, 2 H), 2.80 br s, 2 H), 3.20-3.47 (m, 1 H), 3.62 (d, J=18 Hz, 1 H), 3.82 (br s, 2 H), 3.92 (s, 2 H), 4.38-4.57 (m, 2 H), 5.00-5.05 (m, 1 H), 5.46 (br s, 1 H), 5.57-5.62 (m, 1 H), 7.24 (d, J=8 Hz, 1 H), 7.45-7.48 (n,L, 2 H), 7.60-7.80 (br s, 3 H), 8.21-8.23 (mm, 2 H), 8.70-8.82 (m, 2 H), 9.15 (br s, 1 H), and 9.26 (d, J=8 Hz, 1 H); IR (KBr) 3428, 1772, 1676, 1626, 1202 cm⁻¹ ; MS (ESI) m/e 714 (MH⁺). Anal. Calcd for C₂₈ H₂₉ N₅ O₇ S₃ Cl₂.HCl.CF₃ CO₂ H.0.4 C₄ H₁₀ O: C, 42.42; H, 3.94; N, 7.83. Found: C, 42.16; H, 4.16; N, 7.69.

    __________________________________________________________________________     2.D. Preparation of Additional Representative Thiopyridone Intermediates       The following additional thiopyridone intermediates were prepared              according to the general method of Example                                     2(B) described above:                                                          Intermediate                 .sup.1 H NMR Data         MS                      __________________________________________________________________________                                                            Data                     ##STR39##                                                                                                   ##STR40##                M.sup.+ = 340            ##STR41##                                                                                                   ##STR42##                MH.sup.+  = 369          ##STR43##                                                                                                   ##STR44##                NA                       ##STR45##                                                                                                   ##STR46##                MH.sup.+  = 327          ##STR47##                                                                                                   ##STR48##                (M-H).sup.+ = 353        ##STR49##                                                                                                   ##STR50##                NA                       ##STR51##                                                                                                   ##STR52##                MH.sup.+  = 355          ##STR53##                                                                                                   ##STR54##                NA                       ##STR55##                                                                                                   ##STR56##                NA                       ##STR57##                                                                                                   ##STR58##                MH.sup.+  = 327          ##STR59##                                                                                                   ##STR60##                MH.sup.+  = 355          ##STR61##                                                                                                   ##STR62##                M.sup.+ = 312            ##STR63##                                                                                                   ##STR64##                M.sup.+ = 298            ##STR65##                                                                                                   ##STR66##                (M-H).sup.+ = 339        ##STR67##                                                                                                   ##STR68##                MH.sup.+  = 327          ##STR69##                                                                                                   ##STR70##                M.sup.+ = 268            ##STR71##                                                                                                   ##STR72##                (M-H).sup.+ = 323        ##STR73##                                                                                                   ##STR74##                MH.sup.+  = 440          ##STR75##                                                                                                   ##STR76##                NA                       ##STR77##                                                                                                   ##STR78##                MH.sup.+  = 326          ##STR79##                                                                                                   ##STR80##                MH.sup.+  = 454          ##STR81##                                                                                                   ##STR82##                MH.sup.+  = 440          ##STR83##                                                                                                   ##STR84##                MH.sup.+  = 469          ##STR85##                                                                     __________________________________________________________________________                                   ##STR86##                MH.sup.+  =             __________________________________________________________________________                                                            325                      NA = not available                                                       

The following compounds were prepared according to the general procedures of Examples 1 and 2 by varying the thiol starting material and the pyridine or thiopyridone derivative:

    __________________________________________________________________________      ##STR87##                                                                     Example                                                                        No.   Ar            R.sup.1                 R.sup.2                                                                               R.sup.3                                                                          Isolated                  __________________________________________________________________________                                                          as                         3    2,5-dichlorophenyl                                                                            ##STR88##              H      H CF.sub.3 CO.sub.2 H,                                                           HCl salt                   4    2,5-dichlorophenyl                                                                            ##STR89##              2,6-dimethyl                                                                            CF.sub.3 CO.sub.2 H,                                                           HCl salt                   5    2,5-dichlorophenyl                                                                            ##STR90##              H      H CF.sub.3 CO.sub.2 H,                                                           HCl salt                   6    2,5-dichlorophenyl                                                                            ##STR91##              H      H CF.sub.3 CO.sub.2 H                                                            salt                       7    2,6-dichloropyridin-4-yl                                                                      ##STR92##              H      H CF.sub.3 CO.sub.2 H,                                                           HCl salt                   8    3-bromophenyl                                                                                 ##STR93##              H      H CF.sub.3 CO.sub.2 H,                                                           HCl salt                   9    3,5-dichloro-4-hydroxyphenyl                                                                  ##STR94##              H      H CF.sub.3 CO.sub.2 H                                                            salt                      10    2-chloro-5-trifluoromethylphenyl                                                              ##STR95##              H      H CF.sub.3 CO.sub.2 H,                                                           HCl salt                  11    3,5-dichloro-4-aminophenyl                                                                    ##STR96##              H      H CF.sub.3 CO.sub.2 H                                                            salt                      12    2,5-dimethylphenyl                                                                            ##STR97##              H      H CF.sub.3 CO.sub.2 H,                                                           HCl salt                  13    2,4,5-trichlorophenyl                                                                         ##STR98##              H      H CF.sub.3 CO.sub.2 H,                                                           HCl salt                  14    2,5-dichlorophenyl                                                                            ##STR99##              2,6-dimethyl                                                                            CF.sub.3 CO.sub.2 H                                                            salt                      15    2,6-dichloropyridin-4-yl                                                                      ##STR100##             2,6-dimethyl                                                                            CF.sub.3 CO.sub.2 H                                                            salt                      16                                                                                    ##STR101##                                                                                   ##STR102##             2,6-dimethyl                                                                            HCl salt                  17    3-fluorophenyl                                                                                ##STR103##             2,6-dimethyl                                                                            HCl salt                  18    3-bromophenyl                                                                                 ##STR104##             2,6-dimethyl                                                                             ##STR105##               19    2-chloro-5-trifluoromethylphenyl                                                              ##STR106##             2,6-dimethyl                                                                            CF.sub.3 CO.sub.2 H,                                                           HCl salt                  20    2,4,5-trichlorophenyl                                                                         ##STR107##             2,6-dimethyl                                                                            CF.sub.3 CO.sub.2 H,                                                           HCl salt                  21    2,5-dichlorophenyl                                                                            ##STR108##             2,3-dimethyl                                                                            HCl salt                  22    2,5-dichlorophenyl                                                                            ##STR109##             2-ethyl                                                                               H CF.sub.3 CO.sub.2 H,                                                           HCl salt                  23    2,5-dichlorophenyl                                                                            ##STR110##             2-methyl                                                                              H HCl salt                  24    2,5-dichlorophenyl                                                                            ##STR111##             2-(CH.sub.2).sub.3                                                                    HH.sub.2                                                                         HCl salt                  25    2,5-dichlorophenyl                                                                            ##STR112##             H      H CF.sub.3 CO.sub.2 H                                                            salt                      26                                                                                    ##STR113##                                                                                   ##STR114##             H      H CF.sub.3 CO.sub.2 H                                                            salt                      27    2,5-difluorophenyl                                                                            ##STR115##             2,6-dimethyl                                                                            CF.sub.3 CO.sub.2 H                                                            salt                      28    2,5-dichlorophenyl                                                                            ##STR116##             H      H CF.sub.3 CO.sub.2 H,                                                           HCl salt                  29    2,5-dichlorophenyl                                                                            ##STR117##             H      H CF.sub.3 CO.sub.2 H                                                            salt                      30    2,5-dichlorophenyl                                                                            ##STR118##             H      H CF.sub.3 CO.sub.2 H,                                                           HCl salt                  31    2,5-dichlorophenyl                                                                            ##STR119##             H      H CF.sub.3 CO.sub.2 H,                                                           HCl salt                  32    2,5-dichlorophenyl                                                                            ##STR120##             H      H HCl salt                  33    2,5-dichlorophenyl                                                                            ##STR121##             H      H CF.sub.3 CO.sub.2 H,                                                           HCl salt                  34    2,5-dichlorophenyl                                                                            ##STR122##             H      H HCl salt                  35    2,5-dichlorophenyl                                                                            ##STR123##             H      H HCl salt                  36    2,5-dichlorophenyl                                                                            ##STR124##             H      H CF.sub.3 CO.sub.2 H,                                                           HCl salt                  37    2,5-dichlorophenyl                                                                            ##STR125##             H      H CF.sub.3 CO.sub.2 H,                                                           HCl salt                  38    2,5-dichlorophenyl                                                                            ##STR126##             H      H CF.sub.3 CO.sub.2 H,                                                           HBr salt                  39    2,5-dichlorophenyl                                                                            ##STR127##             H      H CF.sub.3 CO.sub.2 H,                                                           HBr salt                  40    3-iodophenyl                                                                                  ##STR128##             H      H CF.sub.3 CO.sub.2 H                                                            salt                      41    3-chlorophenyl                                                                                ##STR129##             H      H CF.sub.3 CO.sub.2 H                                                            salt                      42    3-iodophenyl                                                                                  ##STR130##             H      H CF.sub.3 CO.sub.2 H                                                            salt                      43    2,5-dichlorophenyl                                                                            ##STR131##             H      H CF.sub.3 CO.sub.2 H                                                            salt                      44    2,5-dichlorophenyl                                                                            ##STR132##             H      H HCl salt                  45    2,5-dichlorophenyl                                                                            ##STR133##             H      H                                                                                 ##STR134##               __________________________________________________________________________

                  TABLE                                                            ______________________________________                                         MS DATA                                                                        Compound of               Method for                                           Example No.    MS          IV to  I                                            ______________________________________                                         1              M.sup.+ = 714                                                                             1                                                    2              MH.sup.+ = 714                                                                            2                                                    3              M.sup.+ = 671                                                                             2                                                    4              MH.sup.+ = 699                                                                            2                                                    5              M.sup.+ = 671                                                                             2                                                    6              MH.sup.+ = 657                                                                            2                                                    7              MH.sup.+ = 658                                                                            2                                                    8              MH.sup.+ = 667                                                                            2                                                    9              M.sup.+ = 673                                                                             2                                                    10             MH.sup.+ = 691                                                                            2                                                    11             MH.sup.+ = 672                                                                            2                                                    12             MH.sup.+ = 617                                                                            2                                                    13             MH.sup.+ = 691                                                                            2                                                    14             MH.sup.+ = 685                                                                            2                                                    15             MH.sup.+ = 686                                                                            2                                                    16             (M-H).sup.+ = 715                                                                         2                                                    17             M.sup.+ = 635                                                                             2                                                    18             MH.sup.+ = 695                                                                            2                                                    19             MH.sup.+ = 719                                                                            2                                                    20             MH.sup.+ = 719                                                                            2                                                    21             MH.sup.+ = 685                                                                            2                                                    22             MH.sup.+ = 685                                                                            2                                                    23             M.sup.+ = 671                                                                             2                                                    24             M.sup.+ = 714                                                                             2                                                    25             M.sup.+ = 657                                                                             2                                                    26             MH.sup.+ = 687                                                                            2                                                    27             MH.sup.+ = 683                                                                            2                                                    28             M.sup.+ = 643                                                                             2                                                    29             MH.sup.+ = 629                                                                            2                                                    30             M.sup.+ = 671                                                                             2                                                    31             M.sup.+ = 657                                                                             2                                                    32             MH.sup.+ = 699                                                                            2                                                    33             M.sup.+ = 657                                                                             2                                                    34             M.sup.+ = 756                                                                             2                                                    35             M.sup.+ = 756                                                                             1                                                    36             M.sup.+ = 728                                                                             2                                                    37             M.sup.+ = 714                                                                             2                                                    38             M.sup.+ = 700                                                                             1                                                    39             M.sup.+ = 742                                                                             1                                                    40             MH+ = 715  2                                                    41             MH+ = 623  2                                                    42             MH+ = 715  2                                                    43             MH+ = 699  2                                                    44             MH+ = 757  2                                                    45             MH+ = 655  2                                                    ______________________________________                                    

                                      TABLE                                        __________________________________________________________________________     NMR DATA                                                                        ##STR135##                                                                    Comp.                                                                          of Ex.                                                                         No. H-2    H-6    H-7    H-3'   H-7'                                                                               NH    ArH    S-PyrH  R                     __________________________________________________________________________      1  3.69 (d, J=18)                                                                        5.12 (d, J=5)                                                                         5.67 (dd, J=                                                                          4.26-4.50 (m,                                                                         3.89 (s)                                                                           9.29 (d, J=8)                                                                        7.22 (dd, J=                                                                          8.00 (m)                                                                               1.63-2.05                 3.80 (d, J=18)                                                                               5,8)   overlaps with    2,8)   8.55 (m)                                                                               (m)                                            R)               7.37-7.58 (m)  2.55-3.05                                                                      (m)                                                                            3.26-3.66                                                                      (m)                                                                            4.00-4.38                                                                      (m, overlaps                                                                   with H-3')                                                                     5.42 (s)                                                                       5.60 (s)               2  3.20-3.47 (m)                                                                         5.00-5.05 (m)                                                                         5.57-5.62 (m)                                                                         4.38-4.57 (m)                                                                         3.92 (s)                                                                           9.26 (d, J=8)                                                                        7.24 (d, J=8)                                                                         8.21-8.23                                                                              1.32-1.57                 3.62 (d, J=18)                        7.45-7.48 (m)                                                                         8.70-8.82                                                                              (m)                                                                            2.27 (br s)                                                                    2.80 (br s)                                                                    3.82 (br s)                                                                    5.46 (br s)                                                                    7.60-7.80                                                                      (m)                                                                            9.15 (br s)            3  3.40 (d, J=17)                                                                        4.99 (d, J=5)                                                                         5.55 (dd, J=                                                                          4.35-4.58 (m)                                                                         3.92 (s)                                                                           9.30 (d, J=8)                                                                        7.22 (dd, J=                                                                          8.06 (d,                                                                               1.04-1.40                 3.59 (d, J=17)                                                                               5,8)   4.56 (d, J=13)   2,8)   8.74 (d,                                                                               (m))                                                            7.43-7.48 (m)  1.77-1.85                                                                      (m)                                                                            3.56 (br s)                                                                    4.43 (br s)            4  3.43 (d, J=17)                                                                        5.04 (d, J=5)                                                                         5.58-5.61 (m)                                                                         4.49 (d, J=13)                                                                        3.94 (s)                                                                           9.29 (d, J=8)                                                                        7.25 (d, J=8)                                                                         2.74 (s,                                                                               1.53 (br s)               3.60-3.66 (m,        4.31-4.52 (m,    7.46-7.49 (m)                                                                         7.87 (s)                                                                               1.79 (br s)               overlaps with        overlaps with                   3.60-3.77                 R)                   R)                              (m, overlaps                                                                   with H-2)                                                                      4.31-4.52                                                                      (m, overlaps                                                                   with H-3')             5  3.47 (d, J=17)                                                                        5.07 (d, J=3)                                                                         5.57-5.62 (m)                                                                         4.34-4.52 (m),                                                                        3.94 (s)                                                                           9.38 (d, J=8)                                                                        7.23 (dd, J=                                                                          8.10 (d,                                                                               1.36-1.39                 3.69 (d, J=17)       overlaps with    2,8)   8.85 (d,                                                                               (m))                                           R)               7.44-7.47 (m)  1.79 (br s)                                                                    4.34-4.52                                                                      (m, overlaps                                                                   with H-3')                                                                     8.38 (br s)            6  3.47 (d, J=17)                                                                        5.09 (d, J=4)                                                                         5.64 (dd, J=                                                                          4.35-4.51 (m,                                                                         3.92 (s)                                                                           9.29 (d, J=8)                                                                        7.23 (dd, J=                                                                          8.06 (d,                                                                               1.75 (br s)               3.69 (d, J=17)                                                                               4,8)   overlaps with    2,8)   8.77 (d,                                                                               1.95 (br s)               overlaps with        R)               7.44-7.74 (m)  3.63-3.75                 R)                                                   (m, overlaps                                                                   with H-2)                                                                      4.35-4.51                                                                      (m, overlaps                                                                   with H-3')             7  3.42 (d, J=17)                                                                        5.02 (d, J=5)                                                                         5.57 (dd, J=                                                                          4.37-4.58 (m,                                                                         3.99 (d,                                                                           9.42 (d, J=8)                                                                        7.51 (s)                                                                              8.10 (d,                                                                               1.72 (br s)               3.61 (d, J=17)                                                                               5,8)   overlaps with                                                                         J=15)            8.73 (d,                                                                               1.98 (br s)                                    R)     4.07 (d,                 3.58-3.74                                             J=15)                    (m)                                                                            4.37-4.58                                                                      (m, overlaps                                                                   with H-3')             8  3.43 (d, J=18)                                                                        5.01 (d, J=5)                                                                         5.56 (dd, J=                                                                          4.30-4.57 (m,                                                                         3.78 (d,                                                                           9.22 (d, J=8)                                                                        7.23 (t, J=7)                                                                         8.09 (br                                                                               1.73 (br s)               3.62 (d, J=18)                                                                               5,8)   overlaps with                                                                         J=15)     7.33 (t, J=7)                                                                         8.75 (br                                                                               2.02 (br s)                                    R)     3.85 (d,  7.55 (d, J=2)  3.55-3.75                                             J=15)                    (m)                                                                            4.30-4.57                                                                      (m, overlaps                                                                   with H-3')             9  3.40 (d, J=18)                                                                        4.96 (d, J=5)                                                                         5.50 (dd, J=                                                                          4.35-4.55 (m,                                                                         4.68 (s)                                                                           9.04 (d, J=8)                                                                        7.39 (s)                                                                              8.14 (d,                                                                               1.65 (br s)               3.61 (d, J=18)                                                                               5.8)   overlaps with           8.73 (d,                                                                               1.90-2.00                                      R)                              (m)                                                                            3.60-3.80                                                                      (m)                                                                            4.35-4.55                                                                      (m, overlaps                                                                   with H-3')            10  3.41 (d, J=18)                                                                        5.00 (d, J=5)                                                                         5.55 (dd, J=                                                                          4.38 (d, J=14)                                                                        3.99 (s)                                                                           9.32 (d, J=8)                                                                        7.53 (d, J=8)                                                                         8.11 (d,                                                                               1.69 (br s)               3.59 (d, J=18)                                                                               5,8)   4.55 (d, J=14)   7.68 (d, J=8)                                                                         8.74 (d,                                                                               2.07 (br s)                                                     7.80 (s)       3.56 (br s)                                                                    4.46 (br s)           11  3.44 (d, J=18)                                                                        5.02 (d, J=4)                                                                         5.56 (dd, J=                                                                          4.39 (d, J=11)                                                                        3.59 (s)                                                                           9.00 (d, J=8)                                                                        7.36 (s)                                                                              8.12 (d,                                                                               1.60-1.80                 3.64 (d, J=18)                                                                               4,8)   4.45-4.55 (m,           8.75 (d,                                                                               (m))                                           overlaps with                   1.90-2.10                                      R)                              (m)                                                                            3.39-3.60                                                                      (m)                                                                            4.45-4.55                                                                      (m)                   12  3.44 (d, J=18)                                                                        5.03 (d, J=5)                                                                         5.59 (dd, J=                                                                          4.38 (d, J=14)                                                                        3.73 (s)                                                                           9.17 (d, J=8)                                                                        6.87 (d, J=8)                                                                         8.08 (br                                                                               1.71 (br s)               3.63 (d, J=18)                                                                               5,8)   4.48 (d, J=14)   7.04 (d, J=8)                                                                         8.74 (br                                                                               1.99 (br s)                                                     7.17 (s)       3.60-3.73                                                                      (m)                                                                            4.40-4.54                                                                      (m)                   13  3.41 (d, J=17)                                                                        4.99 (d, J=5)                                                                         5.54 (dd, J=                                                                          4.36-4.59 (m,                                                                         3.91 (d,                                                                           9.32 (d, J=8)                                                                        7.67 (s)                                                                              8.11 (d,                                                                               1.66 (br s)               3.59 (d, J=17)                                                                               5,8)   overlaps with                                                                         J=15)     7.84 (s)                                                                              8.73 (d,                                                                               2.00 (br s)                                    R)     3.97 (d,                 3.56-3.70                                             J=15)                    (m)                                                                            4.36-4.59                                                                      (m, overlaps                                                                   with H-3')            14  3.46 (d, J=18)                                                                        5.10 (d, J=5)                                                                         5.66 (dd, J=                                                                          4.27-4.40 (m,                                                                         3.90 (s)                                                                           9.28 (d, J=8)                                                                        7.23 (dd, J=                                                                          2.72 (s,                                                                               1.70-2.00                 3.70 (d, J=18)                                                                               5,8)   overlaps with    2,8)   7.84 (s)                                                                               (m)                                            R)               7.46 (d, J=8)  3.47-3.60                                      4.45 (d, J=12)   7.47 (d, J=2)  (m)                                                                            4.27-4.40                                                                      (m, overlaps                                                                   with H-3')            15  3.44 (d, J=18)                                                                        5.07 (d, J=5)                                                                         5.63 (dd, J=                                                                          4.23-4.45 (m,                                                                         3.99 (s)                                                                           9.31 (d, J=8)                                                                        7.49 (s)                                                                              2.72 (s,                                                                               1.74-2.03                 3.68 (d, J=18)                                                                               5,8)   overlaps with           7.80 (s)                                                                               (m)                                            R)                              3.82 (br s)                                    4.39 (d, J=12)                  4.23-4.45                                                                      (m, overlaps                                                                   with H-3')            16  3.50 (d, J=18)                                                                        5.18 (d, J=5)                                                                         5.66 (dd, J=                                                                          4.32 (d, J=13)                                                                        3.89 (s)                                                                           9.29 (d, J=8)                                                                        4.47 (s)                                                                              7.83 (s)                                                                               1.83-2.10                 3.74 (d, J=18)                                                                               5,8)   4.39 (d, J=13)   7.46 (s)                                                                              2.78 (br                                                                               (m)                                                             7.51 (s)                                                                              PyrMe)  3.84-3.99                                                                      (m)                                                                            4.36 (br s)                                                                    8.67 (br s)           17  3.47 (d, J=17)                                                                        5.11 (d, J=4)                                                                         5.67 (dd, J=                                                                          4.30-4.40 (m)                                                                         3.76 (d,                                                                           9.26 (d, J=8)                                                                        6.95 (app ddd,                                                                        7.84 (br                                                                               1.83-2.03                               4,8)          J=14)     J=2, 9, 10)                                                                           2.77 (br                                                                               (m)                                                   3.80 (d,  7.13 (app d, J=                                                                       PyrMe)  3.90-3.96                                             J=14)     10)            (m)                                                             7.25 (ddd, J=  4.29-4.39                                                       2,2,7)         (m)                                                             7.32 (app dd,  8.69 (br s)                                                     J=9,14)                              18  3.42 (d, J=18)                                                                        5.03 (d, J=5)                                                                         5.58 (dd, J=                                                                          4.23-4.49 (m,                                                                         3.79 (s)                                                                           9.15 (d, J=8)                                                                        7.22 (t, J=8)                                                                         2.73 (br                                                                               1.84 (br s)               3.62 (d, J=18)                                                                               5,8)   overlaps with    7.33 (t, J=8)                                                                         PyrMe)  3.57-3.78                                      R)               7.55 (s)                                                                              7.83 (s)                                                                               (m)                                                                            4.23-4.49                                                                      (m, overlaps                                                                   with H-3')            19  3.37 (d, J=18)                                                                        4.99 (d, J=5)                                                                         5.55 (dd, J=                                                                          4.20-4.33 (m,                                                                         3.98 (s)                                                                           9.29 (d, J-8)                                                                        7.53 (d, J=8)                                                                         2.72 (br                                                                               1.85 (br s)               3.55 (d, J=18)                                                                               5,8)   overlaps with    7.68 (d, J=8)                                                                         PyrMe)  3.53 (br s)                                    R)               7.80 (s)                                                                              7.90 (s)                                                                               4.20-4.33                                      4.48 (d, J=14)                  (m, overlaps                                                                   with H-3')            20  3.40 (d, J=18)                                                                        5.04 (d, J=5)                                                                         5.59 (dd, J=                                                                          4.25-4.55 (m,                                                                         3.97 (s)                                                                           9.31 (d, J=8)                                                                        7.69 (s)                                                                              2.75 (br                                                                               1.90 (br s)               3.59 (d, J=18)                                                                               5,8)   overlaps with    7.84 (s)                                                                              PyrMe)  3.57-3.74                                      R)                      7.85 (s)                                                                               (m)                                                                            4.25-4.55                                                                      (m, overlaps                                                                   with H-3')            21  3.51 (d, J=18)                                                                        5.16 (d, J=5)                                                                         5.71 (dd, J=                                                                          4.32-4.42 (m,                                                                         3.93 (s)                                                                           9.28 (d, J=8)                                                                        7.23 (dd, J=                                                                          2.35 (s,                                                                               1.80-2.00                 3.76 (d, J=18)                                                                               5,8)   overlaps with    2,8)   2.72 (s,                                                                               (m)Me)                                         R)               7.46 (d, J=8)                                                                         7.85 (d,                                                                               4.32-4.42                                                       7.50 (d, J=2)  (m, overlaps                                                           8.75 (d,                                                                               with H-3')                                                                     4.50-4.60                                                                      (m)                   22  3.44 (d, J=17)                                                                        5.05 (d, J=5)                                                                         5.59 (dd, J=                                                                          4.36-4.60 (m,                                                                         3.96 (s)                                                                           9.30 (d, J=8)                                                                        7.25 (dd, J=                                                                          1.22-1.38                                                                              1.62-2.08                 3.68 (d, J=17)                                                                               5,8)   overlaps with    2,8)   PyrEt)  (m)                                            R)               7.47 (d, J=8)                                                                         2.95-3.15                                                                              3.50-3.68                                                       7.50 (d, J=2)                                                                         PyrEt)  (m)                                                                    7.96 (br                                                                               4.36-4.60                                                              8.00 (br                                                                               (m, overlaps                                                           8.71 (br                                                                               with H-3')            23  3.48 (d, J=18)                                                                        5.14 (d, J=4)                                                                         5.67 (dd, J=                                                                          4.40-4.50 (m,                                                                         3.93 (s)                                                                           9.28 (d, J=8)                                                                        7.24 (dd, J=                                                                          7.88 (d,                                                                               1.80-2.10                 3.72 (d, J=18)                                                                               4,8)   overlaps with    2,8)   7.96 (s)                                                                               (m)                                            R)               7.46 (d, J=8)                                                                         8.82 (d,                                                                               3.82-4.00                                                       7.47 (app s)                                                                          2.76 (br                                                                               (m)                                                                    PyrMe)  4.40-4.50                                                                      (m, overlaps                                                                   with H-3')                                                                     8.18 (br s)                                                                    8.57 (br s)                                                                    8.66 (br s)           24  3.21 (d, J=18)                                                                        5.16 (d, J=4)                                                                         5.72 (dd, J=                                                                          4.42-4.55 (m)                                                                         3.90 (s)                                                                           9.34 (d, J=7)                                                                        7.24 (dd, J=                                                                          1.60-2.10                                                                              1.60-2.10                 3.39 (d, J=18)                                                                               4,8)                    2,8)   SPyr-CH.sub.2 CH.sub.2,                                                                (m, overlaps                                                    7.47 (d, J=8)                                                                         overlaps                                                                               with                                                            7.48 (app s)                                                                          R)      SPyr-                                                                  2.70-2.81                                                                              CH.sub.2                                                                       CH.sub.2)                                                              (m,     2.70-2.81                                                              SPyr-CH.sub.2,                                                                         (m, overlaps                                                           overlaps                                                                               with                                                                   R)      SPyr-CH.sub.2)                                                         2.83-3.00                                                                              3.00-3.24                                                              7.91 (d,                                                                               (m))                                                                   7.99 (s)                                                                               3.90-4.02                                                              8.85 (d,                                                                               (m))                                                                           8.16 (br s)                                                                    8.45 (br s)                                                                    8.56 (br s)                                                                    8.64 (br s)           25  3.41 (d, J=18)                                                                        5.01 (d, J=5)                                                                         5.54 (dd, J=                                                                          4.39 (d, J=14)                                                                        3.93 (s)                                                                           9.34 (d, J=8)                                                                        7.22 (dd, J=                                                                          8.10 (d,                                                                               1.71 (br s)               3.60 (d, J=18)                                                                               5,8)   4.55 (d, J=14)   2,8)   8.76 (d,                                                                               1.90-2.10                                                       7.44-7.48 (m)  (m)                                                                            3.57 (br s)                                                                    4.43 (br s)                                                                    4.35-4.55                                                                      (m)                   26  3.70 (d, J=18)                                                                        5.07 (d, J=5)                                                                         5.61 (dd, J=                                                                          4.38-4.55 (m,                                                                         3.92 (s)                                                                           9.25 (d, J=8)                                                                        7.47 (s)                                                                              8.11 (d,                                                                               1.90-2.10                 Other peak    5,8)   overlaps with    7.51 (s)                                                                              8.77 (d,                                                                               (m))                      obscured             R)                              4.38-4.55                                                                      (m, overlaps                                                                   with H-3')            27  3.38 (d, J=17)                                                                        5.00 (d, J=5)                                                                         5.57 (dd, J=                                                                          4.25 (d, J=14)                                                                        3.92 (s)                                                                           9.28 (d, J=8)                                                                        7.23 (d, J=8)                                                                         2.72 (s,                                                                               1.70 (br s)               3.61 (d, J=17)                                                                               5,8)   4.45 (d, J=14)   7.44-7.48 (m)                                                                         7.79 (s)                                                                               1.84 (br s)                                                                    3.30-3.70                                                                      (m)                                                                            4.30 (br s)           28  3.44-3.74 (m)                                                                         5.02 (s)                                                                              5.56 (d, J=                                                                           4.42 (br s)                                                                           3.93                                                                               9.38 (d, J=8)                                                                        7.21 (dd, J=                                                                          8.05 (br                                                                               2.41 (br s)                             5,8)   4.67 (br s)                                                                           (br s)    2,8)   8.85 (br                                                                               3.44-3.74                                                       7.41-7.47 (m)  (m)                                                                            3.93 (br s)           29  3.46 (d, J=18)                                                                        5.08 (d, J=5)                                                                         5.61-5.65 (m)                                                                         4.42 (br s)                                                                           3.91 (s)                                                                           9.28 (d, J=8)                                                                        7.22 (d, J=7)                                                                         8.04 (d,                                                                               4.11 (br s)               3.68 (d, J=18)                        7.43-7.46 (m)                                                                         8.69 (d,                                                                               4.72-4.95                                                                      (m)                   30  3.47 (d, J=18)                                                                        5.06-5.07 (m,                                                                         5.81 (dd, J=                                                                          4.48 (br s)                                                                           3.95 (s)                                                                           9.28 (d, J=8)                                                                        7.24 (dd, J=                                                                          8.74 (br                                                                               1.11 (br s)               3.66 (d, J=18)                                                                        overlaps with                                                                         5,8)                    2,8)   8.07 (br                                                                               1.30 (br s)                      R)                             7.45-7.51 (m)  1.55 (br s)                                                                    2.25 (br s)                                                                    2.73 (br s)                                                                    5.06-5.07                                                                      (m, overlaps                                                                   with H-6)                                                                      8.19 (br s)           31  3.64 (d, J=18)                                                                        5.05 (br s)                                                                           5.56-5.60 (m)                                                                         4.46 (br s)                                                                           3.94 (s)                                                                           9.34 (br s)                                                                          7.22 (dd, J=                                                                          8.71 (br                                                                               8.28 (br s)               3.46 (d, J=18)                        3,8)   8.06 (br                                                                               5.05 (br s)                                                     7.43-7.49 (m)  2.79 (br s)                                                                    2.33 (br s)                                                                    2.16 (br s                                                                     1.59 (br s)                                                                    1.37 (br s            32  3.49 (d, J=17)                                                                        5.10 (d, J=5)                                                                         5.62 (dd, J=                                                                          4.44 (br s)                                                                           3.92 (s)                                                                           9.29 (d, J=8)                                                                        7.22 (d, J=8)                                                                         8.01 (d,                                                                               8.41 (br s),              3.67-3.75 (m, 5,8)                    7.44-7.48 (m)                                                                         8.79 (d,                                                                               5.25 (br s),              overlaps with                                        3.67-3.75                 R)                                                   (m, overlaps                                                                   with H-2),                                                                     3.11 (br s),                                                                   2.35 (br s),                                                                   2.20 (br s),                                                                   1.78 (br s),                                                                   1.49 (br s),                                                                   1.19 (br s)           33  3.45 (d, J=17)                                                                        5.06 (d, J=5)                                                                         5.60-5.62 (m)                                                                         4.31-4.46 (m,                                                                         3.91-                                                                              9.34 (d, J=8)                                                                        7.23 (d, J=8)                                                                         8.07-8.11                                                                              2.63-2.75                 3.62-3.68 (m,        overlaps with                                                                         3.95      7.44-7.47 (m)                                                                         overlaps                                                                               (m)h                      overlaps with        R)     (m,              R)      3.62-3.68                 R)                          overlaps         8.90-8.91                                                                              (m, overlaps                                          with R)                  with H-2)                                                                      3.91-3.95                                                                      (m, overlaps                                                                   with H-7')                                                                     4.31-4.46                                                                      (m, overlaps                                                                   with H-3')                                                                     5.40 (br s)                                                                    8.07-8.11                                                                      (m, overlaps                                                                   with SPyrH)           34  3.20-3.50 (m)                                                                         4.99 (d, J=5)                                                                         5.47-5.62 (m,                                                                         4.45 (d, J=14)                                                                        3.90 (s)                                                                           9.25 (d, J=8)                                                                        7.23 (d, J=8)                                                                         8.16 (m)                                                                               1.30 (br s)               3.58 (d, J=18)                                                                               overlaps with                                                                         4.61 (d, J=14)   7.44-7.48 (m)                                                                         8.76 (m)                                                                               1.43 (br s)                             R)                                     2.26 (br s)                                                                    3.12 (br s)                                                                    3.66-3.80                                                                      (m)                                                                            5.47-5.62                                                                      (m, overlaps                                                                   with H-7)                                                                      7.48-7.62                                                                      (m)                   35  3.40-3.59 (m,                                                                         4.98 (d, J=5)                                                                         5.52 (dd, J=                                                                          4.92 (d, J=14)                                                                        3.90 (s)                                                                           9.20 (d, J=8)                                                                        7.23 (dd, J=                                                                          8.09 (d,                                                                               1.40-1.73                 overlaps with 5,8)   4.60 (d, J=14)   2,8)   8.53 (d,                                                                               (m))                      R)                                    7.39-7.53 (m)  2.97-3.16                                                                      (m)                                                                            4.13-4.26                                                                      (m)                                                                            4.79 (s)                                                                       5.16-5.40                                                                      (m)                   36  3.47 (d, J=18)                                                                        5.13 (d, J=5)                                                                         5.69 (dd, J=                                                                          4.29-4.53 (m,                                                                         3.89 (s,                                                                           9.28 (d, J=8)                                                                        7.23 (dd, J=                                                                          7.96 (d,                                                                               1.16-1.47                 3.73 (d, J=18)                                                                               5,8)   overlaps with                                                                         overlaps  2,8)   8.68 (d,                                                                               (m))                                           R)     with R)   7.36-7.55 (m)  1.63-1.95                                                                      (m)                                                                            3.70-3.92                                                                      (m)                                                                            4.38 (br s)           37  3.49 (d, J=18)                                                                        5.13 (d, J=5)                                                                         5.69 (dd, J=                                                                          4.42-4.57 (m)                                                                         3.91 (s)                                                                           9.28 (d, J=8)                                                                        7.24 (dd, J=                                                                          8.03 (d,                                                                               1.55-2.13                 3.73 (d, J=18)                                                                               5,8)                    2,8)   8.75 (d,                                                                               (m))                                                            7.37-7.58 (m)  3.73-4.05                                                                      (m)                                                                            4.38 (br s)           38  3.51 (d, J=18)                                                                        5.14 (d, J=5)                                                                         5.69 (dd, J=                                                                          4.24-4.53 (m)                                                                         3.91 (s)                                                                           9.28 (d, J=8)                                                                        7.23 (dd, J=                                                                          8.08 (m)                                                                               2.92-3.05                 3.75 (d, J=18)                                                                               5,8)                    2,8)   8.62 (m)                                                                               (m)                                                             7.37-7.53 (m)  3.08-3.26                                                                      (m)                                                                            4.24-4.53                                                                      (m)                                                                            5.46 (br s)           39  3.51 (d, J=18)                                                                        5.12 (d, J=5)                                                                         5.67 (dd, J=                                                                          4.26-4.53 (m,                                                                         3.90 (s)                                                                           9.26 (d, J=8)                                                                        7.23 (dd, J=                                                                          8.07 (d,                                                                               3.11-3.71                 3.73 (d, J=18)                                                                               5,8)   overlaps with    2,8)   8.57 (d,                                                                               (m, overlaps                                   R)               7.37-7.55 (m)  with H-2))                                                                     4.00-4.50                                                                      (m, overlaps                                                                   with H-3')                                                                     5.46 (s)                                                                       5.65 (s)              40  3.47 (d, J=17)                                                                        5.07 (d, J=5)                                                                         5.63 (dd, J=                                                                          4.38 (s)                                                                              3.78 (s)                                                                           9.17 (d, J=8)                                                                        7.07 (dd, J=                                                                          8.05 (d,                                                                               1.60-1.80                 3.72 (d, J=17)                                                                               5,8)                    7,7)   8.78 (d,                                                                               (m))                                                            7.33 (d, J=7)  3.66 (br s)                                                     7.52 (d, J=7)  4.37-4.50                                                       7.70 (s)       (m)                   41  3.47 (d, J=17)                                                                        5.11 (d, J=5)                                                                         5.64 (dd, J=                                                                          4.38 (s)                                                                              3.76 (s)                                                                           9.18 (d, J=8)                                                                        7.15-7.23 (m)                                                                         8.04 (d,                                                                               1.60-1.80                 3.75 (d, J=17)                                                                               5,8)                    7.23-7.28 (m)                                                                         8.80 (d,                                                                               (m))                                                            7.42 (s)       3.68 (br s)                                                                    4.37-4.51                                                                      (m)                   42  3.45 (d, J=17)                                                                        5.07 (d, J=5)                                                                         5.62 (dd, J=                                                                          4.31-4.44 (m)                                                                         3.77 (s)                                                                           9.17 (d, J=8)                                                                        7.09 (dd, J=                                                                          8.06 (d,                                                                               1.60-1.80                 3.77 (d, J=17)                                                                               5,8)                    7,7)   8.76 (d,                                                                               (m))                                                            7.34 (d, J=7)  3.68 (br s)                                                     7.52 (d, J=7)  4.42-4.54                                                       7.69 (s)       (m)                   43  3.48 (d, J=18)                                                                        5.13 (d, J=5)                                                                         5.69 (dd, J=                                                                          4.37 (s)                                                                              3.91 (s)                                                                           9.28 (d, J=8)                                                                        7.24 (dd, J=                                                                          8.02 (d,                                                                               1.50-1.70                 3.74 (d, J=18)                                                                               5,8)                    2,8)   8.76 (d,                                                                               (m))                                                            7.45-7.48 (m)  1.80-1.95                                                                      (m)                                                                            4.45 (br s)                                                                    7.87 (d, J=9)         44  3.48 (d, J=18)                                                                        5.13 (d, J=5)                                                                         5.69 (dd, J=                                                                          4.38 (br s)                                                                           3.91 (s)                                                                           9.26 (d, J=8)                                                                        7.23 (dd, J=                                                                          8.01 (d,                                                                               1.36 (s)                  3.74 (d, J=18)                                                                               5,8)                    2,9)   8.75 (d,                                                                               1.40-1.80                                                       7.46 (d, J=9)  (m)                                                             7.47 (d, J=2)  1.83-1.97                                                                      (m)                                                                            4.42 (br s)                                                                    7.08 (d, J=8)         45  3.43 (d, J=17)                                                                        5.04 (d, J=5)                                                                         5.59 (dd, J=                                                                          4.43 (br s)                                                                           3.90 (s)                                                                           9.24 (d, J=8)                                                                        7.22 (dd, J=                                                                          8.11 (d,                                                                               2.42 (br s)               3.63 (d, J=17)                                                                               5,8)                    2,8)   8.83 (d,                                                                               2.93 (br s)                                                     7.44-7.46 (m)  4.10 (br s)                                                                    5.43 (br              __________________________________________________________________________                                                              s)                

We claim:
 1. A compound of the formula ##STR136## wherein R¹ is selected from the group consisting of (a) a C₂ -C₁₀ alkyl group substituted by a carboxyl group and a group of the formula ##STR137## in which R⁹ and R¹⁰ are each independently hydrogen or C₁ -C₆ alkyl, said C₂ -C₁₀ alkyl group being optionally interrupted by one or more nitrogen atoms or carbonyl groups; and(b) a heterocyclic amino acid group of the formula ##STR138## R² and R³ are each independently hydrogen, C₁ -C₆ alkyl or amino(C₁ -C₆)alkyl; or a pharmaceutically acceptable salt thereof.
 2. A compound of the formula ##STR139## wherein R¹ is selected from the group consisting of ##STR140## wherein R⁸ is NR⁹ R¹⁰ or ##STR141## in which R⁹ and R¹⁰ are each independently hydrogen or C₁ -C₆ alkyl; m is 0 or 1; n is as defined above; o is 0 or an integer of from 1 to 4; p is 0 or 1; q is 0 or 1, with the proviso that q is 0 only when p is 0; r and s each represent an integer of from 1 to 4; R¹² is C₁ -C₆ alkyl; and R² and R³ are each independently hydrogen, C₁ -C₆ alkyl or amino(C₁ -C₆)alkyl; or a pharmaceutically acceptable salt thereof. 